阅读说明：本技术 一种干细胞废液沉淀脱色方法及其用途 (Stem cell waste liquid precipitation decoloring method and application thereof ) 是由 许昌啟 于 2021-08-11 设计创作，主要内容包括：本发明公开了本发明一种干细胞废液沉淀脱色方法及其用途,有效地解决了；干细胞在培养基和生理盐水及其一系列地操作过程中的颜色及其中存在的杂质,而当下烧伤、疤痕、伤口修复及手术的应用!本发明实质性让干细胞废液充分得到再利用,着重解决了干细胞废液中杂质对其：蛋白,因子,腺泌体作用的影响!通过本发明的沉淀吸附脱色后的废液中的盐分、杂质、颜色都得到了剔除吸附后将干细胞废液变成肌肤细胞的烧伤、冻伤、疤痕、伤口及术后修复、恢复、再生功能性的需要,实现变废为宝的方法!操作方便,应用面广。(The invention discloses a stem cell waste liquid precipitation decoloring method and application thereof, which effectively solve the problems; the color of stem cells in culture medium and saline and their serial manipulations and the impurities present therein, and the application of current burns, scars, wound repair and surgery! The invention substantially recycles the stem cell waste liquid, and emphasizes solving the problems of impurity in the stem cell waste liquid: effect of protein, factor, glandular action! The method for changing stem cell waste liquid into skin cells after eliminating and adsorbing salt, impurities and colors in the waste liquid after precipitation, adsorption and decoloration of the invention meets the requirements of burning, frostbite, scar and wound changing the stem cell waste liquid into skin cells after adsorption and postoperative repair, recovery and regeneration functionality, and realizes turning waste into wealth! Convenient operation and wide application range.)

1. A method for precipitating and decoloring stem cell waste liquid is characterized by comprising the following steps: the following steps are obtained:

the method comprises the following steps: 1.1: the whole process is sterile, and all detection reports and medical institutions are attached to prove the health of stem cell specimens (without toxicity and side effects) and detection reports and proof documents of mycoplasma, pathogens and infectious diseases;

1.2: aseptically collecting samples in the whole process, and detecting and receiving a report form, a responsibility guarantee certificate and a virus detection report form;

1.3: 98% of stem cell waste liquid remained in the preparation and amplification process of the mesenchymal stem cells under the operation of a whole sterile environment is collected by a centrifuge only at 1800rpm/mis to remove cell flocculation membranes and salt;

1.4, 90 percent of stem cell waste liquid is remained in the whole sterile primary cell collection process; concentrating the stem cell waste liquid at 1800rpm/mis by using a centrifugal machine to remove cell flocculation membranes, salt and tissue debris;

1.5: 96% of stem cell waste liquid remained in the whole sterile liquid changing passage process; concentrating the stem cell waste liquid by a centrifuge at 1800rpm/mis to remove cell flocculation membranes and salt stem cell fragments;

1.6: all the remaining waste liquid is aseptically collected and then is stored in a 4-6 refrigerator or freezer;

step two: 2.1: the purchased resin and activated carbon must have the function of decolorization and must be provided by qualified manufacturers;

2.2: the resin must be tested for bacteria and toxicity at a ratio of 1:10 (1 g resin, 10 g ultrapure water without CO2, mixed and taken out, then placed in a bacteria culture medium, planted in a CO2 incubator, CO 25 ℃, and cultured at 37 ℃ for 24H). Post-observation ensures sterility. According to the proportion of 1: 10: resin 1 g, CO 2-free ultrapure water 10 g, and was taken out for rapid virus detection. After the sterile and nontoxic operation is ensured, the operation environment is entered, and then various subsequent operations are carried out;

2.3: the aseptic operation must be done while receiving and performing the detection on the resin: (masks, goggles, gloves, protective clothing, instruments before and after 75% alcohol disinfection operation, table contact surfaces and spaces for disinfection);

2.4: the glass bottle and jar must have medical qualification prerequisite, must have aseptic report file;

2.5: the glass bottle can is not damaged or defected, if the result is generated or generated in the operation process, the whole operation process can be carried out after restarting and sterilizing from the beginning, and the polluted waste liquid is required to be treated immediately and cannot be stored;

step three: 3. 1: simultaneously carrying out 1 percent (namely resin 1: waste liquid 100 (marked according to the content of the waste liquid)) comparison calculation on the resin and the waste liquid in an aseptic operation platform in an aseptic environment;

3.2: the redundant resin and the waste liquid (marked according to the content of the waste liquid) are aseptically stored and prepared for later use, and the phenomenon that the resin and the waste liquid move and move randomly is avoided during the period, so that the product is prevented from being infected by dust particles and bacteria in the environment;

3.3: taking the same sterile glass bottle, calculating the volume according to the scale or the volume, and adding the volume to the sterile glass bottle according to the proportion of 1% (namely, resin 1, waste liquid 100 (marked according to the content of the waste liquid));

3.4: and (3) transferring the matched waste liquid bottles and cans filled with the waste liquid (marked according to the content of the waste liquid) to a 4-8-degree refrigerator for storage or a sterile operation device for standing and storage. Visual changes were observed in the morning and evening every day starting a week; the color starts to change and thus a recording is made. The next day of the second week, the waste started to become reddish-mixed, and was visually observed once a day and recorded. Starting to fade the color on the second day of the third week, namely, visually observing once on two days and recording; the colour started to sparkle over the weekend. The color is glittering and translucent at the beginning of the fourth week, and the record of one week of decoloring and precipitation is followed up at the moment;

3.5: the formation of a completely transparent layer and an impurity layer of different waste liquid (marked according to the content of the waste liquid) can be seen in the fourth period so as to realize obvious comparison, at the moment, a record is also made, and photos are compared and stored in necessary periods;

3.6: the fibers were clear during this week of visual inspection, and after filtration, they were left for 48 hours and were clearly visible in a microscope as tiny floc fibers. For these fibers there are respectively: protein fibers, factor fibers, cytoplasmic fibers;

3.7: and adding activated carbon (90 percent to 96 percent to 98 percent) in the original proportion into the stock solution product filtered from the stem cell waste liquid, wherein the activated carbon 1 and the raw material stock solution product are 100 percent. The activated carbon of the method has the corresponding grade qualification, and the method can be operated, and the activated carbon detection comprises the following steps: according to the proportion of 1:10 (1 g of activated carbon, 10 g of ultrapure water without CO2, mixed and taken out, then placed in a bacterial culture medium, implanted in a CO2 incubator, CO 25 ℃ and subjected to bacterial culture at the temperature of 37 ℃ for 24H). Post-observation ensures sterility. According to the proportion of 1: 10: 1 g of activated carbon and 10 g of ultrapure water without CO2, and taking out for rapid virus detection test. After the sterile and nontoxic operation is ensured, the operation environment is entered, and then various subsequent operations are carried out;

3.8: the initial week was visually observed to be a dark brown color with a slight cloudiness in the middle, and the initial week was recorded once a day, morning and night. The next day of the second week, the waste started to become reddish-mixed, and was visually observed once a day and recorded. Starting to fade the color on the second day of the third week, namely, visually observing once on two days and recording; the colour started to sparkle over the weekend. The color is glittering and translucent at the beginning of the fourth week, and the record of one week of decoloration and precipitation is followed by the record;

3.9: the sample after standing is obviously glittering and translucent under a microscope, and excessive and residual intractable salt in the sample is fully adsorbed in the activated carbon and is wrapped in impurities on the ground;

3.10 under the normal condition that all factors and proteins of the purified essence are protected, after filtering by a precise filter screen, precipitating for 2 weeks, collecting ultrapure essence, removing precipitates, adding 0.1-0.3% of food-grade and pharmaceutical-grade preservative, and storing at a low temperature of 4-6 ℃. The product is suitable for medical and American and medical grade products, and is added and blended at the low temperature of 20-30 ℃ according to the proportion;

3.11: all operating specifications, operating procedures must be performed aseptically and strictly: the mask, goggles, gloves, protective clothing, instrument and table contact surfaces and spaces before and after 75% alcohol disinfection operation ensure artificial fungus implantation. When the articles are scattered on the ground in the operation process, the articles do not need to be picked up by stretching hands, and after the operation is finished, the articles are collected in a waste bag and cleaned out;

3.12: the PH of the raw material stock solution can be adjusted to the compound required degree in the buffer solution; it is generally recommended that between PH6-7.5, more next-step executions can be performed to meet each need.

2. The method for precipitation and decoloration of stem cell waste liquid and the application thereof according to claim 1, wherein the method comprises the following steps: according to scheme 1.3 of step one: 98% of stem cell waste liquid remained in the preparation and amplification process of the mesenchymal stem cells under the operation of a whole sterile environment is collected by a centrifuge only at 1800rpm/mis to remove cell flocculation membranes and salt;

the scheme 1.4 comprises that 90 percent of stem cell waste liquor is remained in the whole sterile primary cell collection process; concentrating the stem cell waste liquid at 1800rpm/mis by using a centrifugal machine to remove cell flocculation membranes, salt and tissue debris;

scheme 1.5: 96% of stem cell waste liquid remained in the whole sterile liquid changing passage process; concentrating the stem cell waste liquid by a centrifuge at 1800rpm/mis to remove cell flocculation membranes and salt stem cell fragments;

scheme 1.6: all remaining waste liquid is aseptically collected and stored in a 4-6 refrigerator or freezer.

3. The method for precipitation and decoloration of stem cell waste liquid and the application thereof according to claim 1, wherein the method comprises the following steps: according to scheme 2.2 described in step two: the resin must be tested for bacteria and toxicity at a ratio of 1:10 (1 g resin, 10 g ultrapure water without CO2, mixed and taken out, then placed in a bacteria culture medium, planted in a CO2 incubator, CO 25 ℃, and cultured at 37 ℃ for 24H). Post-observation ensures sterility. According to the proportion of 1: 10: resin 1 g, CO 2-free ultrapure water 10 g, and was taken out for rapid virus detection. After the sterile and nontoxic operation is ensured, the operation environment is entered, and then various subsequent operations are carried out;

scheme 2.3: the aseptic operation must be done while receiving and performing the detection on the resin: (masks, goggles, gloves, protective clothing, instruments before and after 75% alcohol disinfection operation, table contact surfaces and spaces for disinfection);

scheme 2.5: the glass bottle can is not damaged or defected, if the damage or the defection occurs or occurs in the operation process, the whole operation process can be restarted after the disinfection and the sterilization are started from the beginning, and the polluted waste liquid is required to be treated immediately and is not stored.

4. The surface anti-fingerprint treatment process of the ultrathin magnesium alloy notebook computer shell as claimed in claim 1, characterized in that: according to the scheme 3.1 of the third step, the resin and the waste liquid are subjected to 1 percent (namely, the resin 1: the waste liquid 100 (marked according to the content of the waste liquid)) proportion calculation under the aseptic environment and then are placed in an aseptic operation table;

further, the method comprises the following steps of; scheme 3.2: the redundant resin and the waste liquid (marked according to the content of the waste liquid) are aseptically stored and prepared for later use, and the phenomenon that the resin and the waste liquid move and move randomly is avoided during the period, so that the product is prevented from being infected by dust particles and bacteria in the environment;

further optimization; scheme 3.3: taking the same sterile glass bottle, calculating the volume according to the scale or the volume, and adding the volume to the sterile glass bottle according to the proportion of 1% (namely, resin 1, waste liquid 100 (marked according to the content of the waste liquid));

further optimization; scheme 3.4: and (3) transferring the matched waste liquid bottles and cans filled with the waste liquid (marked according to the content of the waste liquid) to a 4-8-degree refrigerator for storage or a sterile operation device for standing and storage. Visual changes were observed in the morning and evening every day starting a week; the color starts to change and thus a recording is made. The next day of the second week, the waste started to become reddish-mixed, and was visually observed once a day and recorded. Starting to fade the color on the second day of the third week, namely, visually observing once on two days and recording; the colour started to sparkle over the weekend. The color is glittering and translucent at the beginning of the fourth week, and the record of one week of decoloring and precipitation is followed up at the moment;

further optimization; scheme 3.5: the formation of a completely transparent layer and an impurity layer of different waste liquid (marked according to the content of the waste liquid) can be seen in the fourth period so as to realize obvious comparison, at the moment, a record is also made, and photos are compared and stored in necessary periods;

scheme 3.6: the fibers were clear during this week of visual inspection, and after filtration, they were left for 48 hours and were clearly visible in a microscope as tiny floc fibers. For these fibers there are respectively: protein fibers, factor fibers, cytoplasmic fibers;

and 3.2, performing re-decolorization, process operation, process notes and process tracking notes on the purified waste liquid by using the purified activated carbon according to the proportion according to the process flow, proportion, operation specification and notes of activated carbon detection. Aims to ensure that various factors and proteins secreted by stem cells in the purified essence are substantially protected from being damaged by activity, 0.1-0.3% of food and drug grade preservative is added, and the essence is stored at a low temperature of 4-6 ℃. The product is suitable for medical and American and medical grade products;

scheme 3.7: and adding activated carbon (90 percent to 96 percent to 98 percent) in the original proportion into the stock solution product filtered from the stem cell waste liquid, wherein the activated carbon 1 and the raw material stock solution product are 100 percent. The activated carbon of the method has the corresponding grade qualification, and the method can be operated, and the activated carbon detection comprises the following steps: according to the proportion of 1:10 (1 g of activated carbon, 10 g of ultrapure water without CO2, mixed and taken out, then placed in a bacterial culture medium, implanted in a CO2 incubator, CO 25 ℃ and subjected to bacterial culture at the temperature of 37 ℃ for 24H). Post-observation ensures sterility. According to the proportion of 1: 10: 1 g of activated carbon and 10 g of ultrapure water without CO2, and taking out for rapid virus detection test. After the sterile and nontoxic operation is ensured, the operation environment is entered, and then various subsequent operations are carried out;

scheme 3.8: the initial week was visually observed to be a dark brown color with a slight cloudiness in the middle, and the initial week was recorded once a day, morning and night. The next day of the second week, the waste started to become reddish-mixed, and was visually observed once a day and recorded. Starting to fade the color on the second day of the third week, namely, visually observing once on two days and recording; the colour started to sparkle over the weekend. The color is glittering and translucent at the beginning of the fourth week, and the record of one week of decoloration and precipitation is followed by the record;

scheme 3.9: the sample after standing is obviously glittering and translucent under a microscope, and excessive and residual intractable salt in the sample is fully adsorbed in the activated carbon and is wrapped in impurities on the ground;

in the scheme 3.10, under the normal condition that all factors and proteins of the purified essence are protected, the purified essence is filtered by a precise filter screen, then is precipitated for 2 weeks, and is collected with ultrapure essence, the precipitate is removed, 0.1-0.3% of food-grade and pharmaceutical-grade preservative is added, and the mixture is placed at a low temperature of 4-6 ℃ for preservation. The product is suitable for medical and American and medical grade products, and is added and blended at the low temperature of 20-30 ℃ according to the proportion;

scheme 3.11: all operating specifications, operating procedures must be performed aseptically and strictly: the mask, goggles, gloves, protective clothing, instrument and table contact surfaces and spaces before and after 75% alcohol disinfection operation ensure artificial fungus implantation. When the articles are scattered on the ground in the operation process, the articles do not need to be picked up by stretching hands, and after the operation is finished, the articles are collected in a waste bag and cleaned out;

scheme 3.12: the PH of the raw material stock solution can be adjusted to the compound required degree in the buffer solution; it is generally recommended that between PH6-7.5, more next-step executions can be performed to meet each need.

Technical Field

The invention relates to the biotechnology field of recycling stem cell waste liquid resources left after dry cells are finished in GMP space, in particular to a stem cell waste liquid precipitation and decoloration method and application thereof.

Background

The stem cell MSC preparation technology is improved and perfected, wherein a plurality of technologies are accepted by international and domestic expert communities, in Mesenchymal stem cell Mesenchymal stem cells, Mesenchymal stem cell suspension under the MSCs technology acts on the aspect of human health, stem cells have acquired the efficiency of the minister and the minister which can not achieve the purpose, and stem cell waste liquid forgotten in the stem cell preparation process is ignored, so that the biological treasure resource: among them are proteins, factors, adenosomes, and many of them are applied to the skin cells, which require EGF/FGF of biological cells, superoxide dismutase SOD! In the preparation and amplification procedure of the umbilical cord mesenchymal stem cells, a series of strict screens are carried out: starting coating digestion, and whole-course aseptic sealing of the specimen to sample collection and detection: virus detection, disinfection treatment, antibody control after further disinfection treatment, high-temperature sterilization, digestion culture, subculture and cell collection! The waste liquid generated in the series of processes is stored in a sterile environment and collected in a waste liquid tank! The series of steps are performed in a GMP environment, all according to strict rule-bound behavior criteria! The used consumable reagents must have corresponding approved documents, so that the safety of the mesenchymal stem cells is sufficiently guaranteed. The safety of the waste liquid source is not described much, so for the safe treatment and collection of the waste liquid, the waste liquid must be processed in the same (GMP) environment as the mesenchymal stem cell operation environment, and the consumable reagent raw materials for waste liquid treatment also have corresponding approval documents to ensure the implementation of the steps in the waste liquid treatment process! The quality control in all aspects is strict in the waste liquid treatment step, and the safety, effectiveness and practicability of the stem cell waste liquid treatment are ensured.

Disclosure of Invention

The invention aims to provide a method for precipitating and decoloring stem cell waste liquid and application thereof, aiming at solving the defect that the mesenchymal stem cell waste liquid is wasted after neglected in the current operation technology, the invention provides the method for precipitating and decoloring the stem cell waste liquid and the application thereof: the method provides a pure stock solution raw material deficiency supply for skin cell burn, cold injury, scar and wound and postoperative repair, recovery and regeneration functional requirements in future; whether protein or factor is extracted, or SOD is extracted and then is added into preparations and original liquid cream gel required by skin in a proper amount, and the problems of substantial safety and obvious effect are solved.

In order to achieve the purpose, the invention provides the following technical scheme: a method for precipitation and decoloration of stem cell waste liquid comprises the following steps:

the method comprises the following steps: 1.1: the whole process is sterile, and all detection reports and medical institutions are attached to prove the health of stem cell specimens (without toxicity and side effects) and detection reports and proof documents of mycoplasma, pathogens and infectious diseases;

1.2: aseptically collecting samples in the whole process, and detecting and receiving a report form, a responsibility guarantee certificate and a virus detection report form;

1.3: 98% of stem cell waste liquid remained in the preparation and amplification process of the mesenchymal stem cells under the operation of a whole sterile environment is collected by a centrifuge only at 1800rpm/mis to remove cell flocculation membranes and salt;

1.4, 90 percent of stem cell waste liquid is remained in the whole sterile primary cell collection process; concentrating the stem cell waste liquid at 1800rpm/mis by using a centrifugal machine to remove cell flocculation membranes, salt and tissue debris;

1.5: 96% of stem cell waste liquid remained in the whole sterile liquid changing passage process; concentrating the stem cell waste liquid by a centrifuge at 1800rpm/mis to remove cell flocculation membranes and salt stem cell fragments;

1.6: all the remaining waste liquid is aseptically collected and then is stored in a 4-6 refrigerator or freezer;

step two: 2.1: the purchased resin and activated carbon must have the function of decolorization and must be provided by qualified manufacturers;

2.2: the resin must be tested for bacteria and toxicity at a ratio of 1:10 (1 g resin, 10 g ultrapure water without CO2, mixed and taken out, then placed in a bacteria culture medium, planted in a CO2 incubator, CO 25 ℃, and cultured at 37 ℃ for 24H). Post-observation ensures sterility. According to the proportion of 1: 10: resin 1 g, CO 2-free ultrapure water 10 g, and was taken out for rapid virus detection. After the sterile and nontoxic operation is ensured, the operation environment is entered, and then various subsequent operations are carried out;

2.3: the aseptic operation must be done while receiving and performing the detection on the resin: (masks, goggles, gloves, protective clothing, instruments before and after 75% alcohol disinfection operation, table contact surfaces and spaces for disinfection);

2.4: the glass bottle and jar must have medical qualification prerequisite, must have aseptic report file;

2.5: the glass bottle can is not damaged or defected, if the result is generated or generated in the operation process, the whole operation process can be carried out after restarting and sterilizing from the beginning, and the polluted waste liquid is required to be treated immediately and cannot be stored;

step three: 3. 1: simultaneously carrying out 1 percent (namely resin 1: waste liquid 100 (marked according to the content of the waste liquid)) comparison calculation on the resin and the waste liquid in an aseptic operation platform in an aseptic environment;

3.2: the redundant resin and the waste liquid (marked according to the content of the waste liquid) are aseptically stored and prepared for later use, and the phenomenon that the resin and the waste liquid move and move randomly is avoided during the period, so that the product is prevented from being infected by dust particles and bacteria in the environment;

3.3: taking the same sterile glass bottle, calculating the volume according to the scale or the volume, and adding the volume to the sterile glass bottle according to the proportion of 1% (namely, resin 1, waste liquid 100 (marked according to the content of the waste liquid));

3.4: and (3) transferring the matched waste liquid bottles and cans filled with the waste liquid (marked according to the content of the waste liquid) to a 4-8-degree refrigerator for storage or a sterile operation device for standing and storage. Visual changes were observed in the morning and evening every day starting a week; the color starts to change and thus a recording is made. The next day of the second week, the waste started to become reddish-mixed, and was visually observed once a day and recorded. Starting to fade the color on the second day of the third week, namely, visually observing once on two days and recording; the colour started to sparkle over the weekend. The color is glittering and translucent at the beginning of the fourth week, and the record of one week of decoloring and precipitation is followed up at the moment;

3.5: the formation of a completely transparent layer and an impurity layer of different waste liquid (marked according to the content of the waste liquid) can be seen in the fourth period so as to realize obvious comparison, at the moment, a record is also made, and photos are compared and stored in necessary periods;

3.6: the fibers were clear during this week of visual inspection, and after filtration, they were left for 48 hours and were clearly visible in a microscope as tiny floc fibers. For these fibers there are respectively: protein fibers, factor fibers, cytoplasmic fibers;

3.7: and adding activated carbon (90 percent to 96 percent to 98 percent) in the original proportion into the stock solution product filtered from the stem cell waste liquid, wherein the activated carbon 1 and the raw material stock solution product are 100 percent. The activated carbon of the method has the corresponding grade qualification, and the method can be operated, and the activated carbon detection comprises the following steps: according to the proportion of 1:10 (1 g of activated carbon, 10 g of ultrapure water without CO2, mixed and taken out, then placed in a bacterial culture medium, implanted in a CO2 incubator, CO 25 ℃ and subjected to bacterial culture at the temperature of 37 ℃ for 24H). Post-observation ensures sterility. According to the proportion of 1: 10: 1 g of activated carbon and 10 g of ultrapure water without CO2, and taking out for rapid virus detection test. After the sterile and nontoxic operation is ensured, the operation environment is entered, and then various subsequent operations are carried out;

3.8: the initial week was visually observed to be a dark brown color with a slight cloudiness in the middle, and the initial week was recorded once a day, morning and night. The next day of the second week, the waste started to become reddish-mixed, and was visually observed once a day and recorded. Starting to fade the color on the second day of the third week, namely, visually observing once on two days and recording; the colour started to sparkle over the weekend. The color is glittering and translucent at the beginning of the fourth week, and the record of one week of decoloration and precipitation is followed by the record;

3.9: the sample after standing is obviously glittering and translucent under a microscope, and excessive and residual intractable salt in the sample is fully adsorbed in the activated carbon and is wrapped in impurities on the ground;

3.10 under the normal condition that all factors and proteins of the purified essence are protected, after filtering by a precise filter screen, precipitating for 2 weeks, collecting ultrapure essence, removing precipitates, adding 0.1-0.3% of food-grade and pharmaceutical-grade preservative, and storing at a low temperature of 4-6 ℃. The product is suitable for medical and American and medical grade products, and is added and blended at the low temperature of 20-30 ℃ according to the proportion;

3.11: all operating specifications, operating procedures must be performed aseptically and strictly: the mask, goggles, gloves, protective clothing, instrument and table contact surfaces and spaces before and after 75% alcohol disinfection operation ensure artificial fungus implantation. When the articles are scattered on the ground in the operation process, the articles do not need to be picked up by stretching hands, and after the operation is finished, the articles are collected in a waste bag and cleaned out;

3.12: the PH of the raw material stock solution can be adjusted to the compound required degree in the buffer solution; it is generally recommended that between PH6-7.5, more next-step executions can be performed to meet each need.

Preferably, according to step one, scheme 1.3: 98% of stem cell waste liquid remained in the preparation and amplification process of the mesenchymal stem cells under the operation of a whole sterile environment is collected by a centrifuge only at 1800rpm/mis to remove cell flocculation membranes and salt;

the scheme 1.4 comprises that 90 percent of stem cell waste liquor is remained in the whole sterile primary cell collection process; concentrating the stem cell waste liquid at 1800rpm/mis by using a centrifugal machine to remove cell flocculation membranes, salt and tissue debris;

scheme 1.5: 96% of stem cell waste liquid remained in the whole sterile liquid changing passage process; concentrating the stem cell waste liquid by a centrifuge at 1800rpm/mis to remove cell flocculation membranes and salt stem cell fragments;

scheme 1.6: all remaining waste liquid is aseptically collected and stored in a 4-6 refrigerator or freezer.

Preferably, according to scheme 2.2 of step two: the resin must be tested for bacteria and toxicity at a ratio of 1:10 (1 g resin, 10 g ultrapure water without CO2, mixed and taken out, then placed in a bacteria culture medium, planted in a CO2 incubator, CO 25 ℃, and cultured at 37 ℃ for 24H). Post-observation ensures sterility. According to the proportion of 1: 10: resin 1 g, CO 2-free ultrapure water 10 g, and was taken out for rapid virus detection. After the sterile and nontoxic operation is ensured, the operation environment is entered, and then various subsequent operations are carried out;

scheme 2.3: the aseptic operation must be done while receiving and performing the detection on the resin: (masks, goggles, gloves, protective clothing, instruments before and after 75% alcohol disinfection operation, table contact surfaces and spaces for disinfection);

scheme 2.5: the glass bottle can is not damaged or defected, if the damage or the defection occurs or occurs in the operation process, the whole operation process can be restarted after the disinfection and the sterilization are started from the beginning, and the polluted waste liquid is required to be treated immediately and is not stored.

According to the scheme 3.1 of the third step, the resin and the waste liquid are subjected to 1 percent (namely, the resin 1: the waste liquid 100 (marked according to the content of the waste liquid)) proportion calculation under the aseptic environment and then are placed in an aseptic operation table;

further, the method comprises the following steps of; scheme 3.2: the redundant resin and the waste liquid (marked according to the content of the waste liquid) are aseptically stored and prepared for later use, and the phenomenon that the resin and the waste liquid move and move randomly is avoided during the period, so that the product is prevented from being infected by dust particles and bacteria in the environment;

further optimization; scheme 3.3: taking the same sterile glass bottle, calculating the volume according to the scale or the volume, and adding the volume to the sterile glass bottle according to the proportion of 1% (namely, resin 1, waste liquid 100 (marked according to the content of the waste liquid));

further optimization; scheme 3.4: and (3) transferring the matched waste liquid bottles and cans filled with the waste liquid (marked according to the content of the waste liquid) to a 4-8-degree refrigerator for storage or a sterile operation device for standing and storage. Visual changes were observed in the morning and evening every day starting a week; the color starts to change and thus a recording is made. The next day of the second week, the waste started to become reddish-mixed, and was visually observed once a day and recorded. Starting to fade the color on the second day of the third week, namely, visually observing once on two days and recording; the colour started to sparkle over the weekend. The color is glittering and translucent at the beginning of the fourth week, and the record of one week of decoloring and precipitation is followed up at the moment;

further optimization; scheme 3.5: the formation of a completely transparent layer and an impurity layer of different waste liquid (marked according to the content of the waste liquid) can be seen in the fourth period so as to realize obvious comparison, at the moment, a record is also made, and photos are compared and stored in necessary periods;

scheme 3.6: the fibers were clear during this week of visual inspection, and after filtration, they were left for 48 hours and were clearly visible in a microscope as tiny floc fibers. For these fibers there are respectively: protein fibers, factor fibers, cytoplasmic fibers;

and 3.2, performing re-decolorization, process operation, process notes and process tracking notes on the purified waste liquid by using the purified activated carbon according to the proportion according to the process flow, proportion, operation specification and notes of activated carbon detection. Aims to ensure that various factors and proteins secreted by stem cells in the purified essence are substantially protected from being damaged by activity, 0.1-0.3% of food and drug grade preservative is added, and the essence is stored at a low temperature of 4-6 ℃. The product is suitable for medical and American and medical grade products;

scheme 3.7: and adding activated carbon (90 percent to 96 percent to 98 percent) in the original proportion into the stock solution product filtered from the stem cell waste liquid, wherein the activated carbon 1 and the raw material stock solution product are 100 percent. The activated carbon of the method has the corresponding grade qualification, and the method can be operated, and the activated carbon detection comprises the following steps: according to the proportion of 1:10 (1 g of activated carbon, 10 g of ultrapure water without CO2, mixed and taken out, then placed in a bacterial culture medium, implanted in a CO2 incubator, CO 25 ℃ and subjected to bacterial culture at the temperature of 37 ℃ for 24H). Post-observation ensures sterility. According to the proportion of 1: 10: 1 g of activated carbon and 10 g of ultrapure water without CO2, and taking out for rapid virus detection test. After the sterile and nontoxic operation is ensured, the operation environment is entered, and then various subsequent operations are carried out;

scheme 3.8: the initial week was visually observed to be a dark brown color with a slight cloudiness in the middle, and the initial week was recorded once a day, morning and night. The next day of the second week, the waste started to become reddish-mixed, and was visually observed once a day and recorded. Starting to fade the color on the second day of the third week, namely, visually observing once on two days and recording; the colour started to sparkle over the weekend. The color is glittering and translucent at the beginning of the fourth week, and the record of one week of decoloration and precipitation is followed by the record;

scheme 3.9: the sample after standing is obviously glittering and translucent under a microscope, and excessive and residual intractable salt in the sample is fully adsorbed in the activated carbon and is wrapped in impurities on the ground;

in the scheme 3.10, under the normal condition that all factors and proteins of the purified essence are protected, the purified essence is filtered by a precise filter screen, then is precipitated for 2 weeks, and is collected with ultrapure essence, the precipitate is removed, 0.1-0.3% of food-grade and pharmaceutical-grade preservative is added, and the mixture is placed at a low temperature of 4-6 ℃ for preservation. The product is suitable for medical and American and medical grade products, and is added and blended at the low temperature of 20-30 ℃ according to the proportion;

scheme 3.11: all operating specifications, operating procedures must be performed aseptically and strictly: the mask, goggles, gloves, protective clothing, instrument and table contact surfaces and spaces before and after 75% alcohol disinfection operation ensure artificial fungus implantation. When the articles are scattered on the ground in the operation process, the articles do not need to be picked up by stretching hands, and after the operation is finished, the articles are collected in a waste bag and cleaned out;

scheme 3.12: the PH of the raw material stock solution can be adjusted to the compound required degree in the buffer solution; it is generally recommended that between PH6-7.5, more next executions can be executed according to each need.

Compared with the prior art, the invention has the beneficial effects that:

the color of stem cells in culture medium and saline and their serial manipulations and the impurities present therein, and the application of current burns, scars, wound repair and surgery! The invention substantially recycles the stem cell waste liquid, and emphasizes solving the problems of impurity in the stem cell waste liquid: effect of protein, factor, glandular action! The method for changing the stem cell waste liquid into skin cells after eliminating and adsorbing salt, impurities and colors in the waste liquid after precipitation, adsorption and decoloration of the invention meets the requirements of burn, frostbite, scar and wound changing the stem cell waste liquid into skin cells after adsorption and the functions of repair, recovery and regeneration after operation, realizes the method for changing waste into valuable, has convenient operation and wide application range

Detailed Description

The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

A surface anti-fingerprint treatment process of an ultrathin magnesium alloy notebook computer shell comprises the following steps:

scheme 1:

1.1: the whole process is sterile, and all detection reports and medical institutions are attached to prove the health (no toxic or side effect) of stem cell specimens and the detection reports and the proof documents of mycoplasma, pathogens and infectious diseases.

1.2: sterile sampling is carried out in the whole process, and a detection acceptance report sheet, a responsibility guarantee certificate and a virus detection report sheet are obtained.

1.3: the stem cell waste liquid which is left in the preparation and amplification procedure of the mesenchymal stem cells under the operation of the whole sterile environment is 98 percent, and the stem cell waste liquid is concentrated by a centrifuge at 1800rpm/mis to remove cell flocculation membranes and salt.

1.4, 90 percent of stem cell waste liquid is remained in the whole sterile primary cell collection process; and (4) concentrating the stem cell waste liquid by a centrifuge at 1800rpm/mis to remove cell flocculation membranes, salt and tissue debris.

1.5: 96% of stem cell waste liquid remained in the whole sterile liquid changing passage process; and (4) concentrating the stem cell waste liquid by a centrifuge at 1800rpm/mis to remove cell flocculation membranes and salt stem cell fragments.

1.6: all remaining waste liquid is aseptically collected and stored in a 4-6 refrigerator or freezer.

Scheme 2:

2.1: the purchased resin and activated carbon must have the function of decolorization and must be provided by qualified manufacturers.

2.2: the resin must be tested for bacteria and toxicity at a ratio of 1:10 (1 g resin, 10 g ultrapure water without CO2, mixed and taken out, then placed in a bacteria culture medium, planted in a CO2 incubator, CO 25 ℃, and cultured at 37 ℃ for 24H). Post-observation ensures sterility. According to the proportion of 1: 10: resin 1 g, CO 2-free ultrapure water 10 g, and was taken out for rapid virus detection. After the sterile and nontoxic operation is ensured, the operation environment is entered, and then various subsequent operations are carried out.

2.3: the aseptic operation must be done while receiving and performing the detection on the resin: (masks, goggles, gloves, protective clothing, instruments before and after 75% alcohol disinfection operation, table contact surfaces and spaces).

2.4: the glass bottle and jar must have medical qualification, must have a sterile report file,

2.5: the glass bottle can is not damaged or defected, if the damage or the defection occurs or occurs in the operation process, the whole operation process can be restarted after the disinfection and the sterilization are started from the beginning, and the polluted waste liquid is required to be treated immediately and is not stored.

Scheme 3:

3.1: the resin and waste liquid are subjected to 1% (namely, resin 1: waste liquid 100 (marked according to the content of the waste liquid)) ratio calculation under a sterile environment and then are placed in a sterile operation table.

3.2: the redundant resin and the waste liquid (marked according to the content of the waste liquid) are aseptically stored and are ready for later use, and the phenomenon that the resin and the waste liquid move randomly and do not move randomly is avoided during the period, so that the product is prevented from being infected by dust particles and bacteria in the environment.

3.3: the same sterile glass bottle is taken, and the volume is calculated according to the scale or the volume and then added according to the proportion of 1 percent (namely, the resin 1 and the waste liquid 100 (marked according to the content of the waste liquid)) into the sterile glass bottle.

3.4: and (3) transferring the matched waste liquid bottles and cans filled with the waste liquid (marked according to the content of the waste liquid) to a 4-8-degree refrigerator for storage or a sterile operation device for standing and storage. Visual changes were observed in the morning and evening every day starting a week; the color starts to change and thus a recording is made. The next day of the second week, the waste started to become reddish-mixed, and was visually observed once a day and recorded. Starting to fade the color on the second day of the third week, namely, visually observing once on two days and recording; the colour started to sparkle over the weekend. The colour was clear at the beginning of the fourth week, and a record of one week was made during this time with the colour removal precipitation.

3.5: the complete transparent layer and the impurity layer of the waste liquid with different contents (marked according to the content of the waste liquid) can be seen to form obvious contrast in the fourth period, the record is also made at the moment, and the picture is compared and stored at the necessary period.

3.6: the fibers were clear during this week of visual inspection, and after filtration, they were left for 48 hours and were clearly visible in a microscope as tiny floc fibers. For these fibers there are respectively: protein fibers, factor fibers, cytoplasmic fibers.

3.7: and adding activated carbon (90 percent to 96 percent to 98 percent) in the original proportion into the stock solution product filtered from the stem cell waste liquid, wherein the activated carbon 1 and the raw material stock solution product are 100 percent. The activated carbon of the method has the corresponding grade qualification, and the method can be operated, and the activated carbon detection comprises the following steps: according to the proportion of 1:10 (1 g of activated carbon, 10 g of ultrapure water without CO2, mixed and taken out, then placed in a bacterial culture medium, implanted in a CO2 incubator, CO 25 ℃ and subjected to bacterial culture at the temperature of 37 ℃ for 24H). Post-observation ensures sterility. According to the proportion of 1: 10: 1 g of activated carbon and 10 g of ultrapure water without CO2, and taking out for rapid virus detection test. After the sterile and nontoxic operation is ensured, the operation environment is entered, and then various subsequent operations are carried out.

3.8: the initial week was visually observed to be a dark brown color with a slight cloudiness in the middle, and the initial week was recorded once a day, morning and night. The next day of the second week, the waste started to become reddish-mixed, and was visually observed once a day and recorded. Starting to fade the color on the second day of the third week, namely, visually observing once on two days and recording; the colour started to sparkle over the weekend. The color was clear at the beginning of the fourth week, and a record of one week was made during this time with the decolorizing precipitation.

3.9: the sample after standing is obviously glittering and translucent under a microscope, and the excessive and residual intractable salt in the sample is fully adsorbed in the activated carbon and wrapped in the impurities on the ground.

3.10 under the normal condition that all factors and proteins of the purified essence are protected, after filtering by a precise filter screen, precipitating for 2 weeks, collecting ultrapure essence, removing precipitates, adding 0.1-0.3% of food-grade and pharmaceutical-grade preservative, and storing at a low temperature of 4-6 ℃. The product is suitable for medical and American products and medical grade products, and is added and blended at the low temperature of 20-30 ℃ according to the proportion.

3.11: all operating specifications, operating procedures must be performed aseptically and strictly: the mask, goggles, gloves, protective clothing, instrument and table contact surfaces and spaces before and after 75% alcohol disinfection operation ensure artificial fungus implantation. When the articles are scattered on the ground in the operation process, the articles do not need to be picked up by stretching hands, and after the operation is finished, the articles are collected in the waste bag and are cleaned out.

3.12: the PH of the raw material stock solution can be adjusted to the compound required degree in the buffer solution; it is generally recommended that between PH6-7.5, more next-step executions can be performed to meet each need.

The method comprises the following steps: mainly aims at the test procedures of different waste liquid sources and aims at further purification of decoloration and execution of the requirement of preservation environment.

Further, the method comprises the following steps of; scheme 1.3: the stem cell waste liquid which is left in the preparation and amplification procedure of the mesenchymal stem cells under the operation of the whole sterile environment is 98 percent, and the stem cell waste liquid is concentrated by a centrifuge at 1800rpm/mis to remove cell flocculation membranes and salt.

Further, the method comprises the following steps of; the scheme 1.4 comprises that 90 percent of stem cell waste liquor is remained in the whole sterile primary cell collection process; and (4) concentrating the stem cell waste liquid by a centrifuge at 1800rpm/mis to remove cell flocculation membranes, salt and tissue debris.

Further optimization; scheme 1.5: 96% of stem cell waste liquid remained in the whole sterile liquid changing passage process; and (4) concentrating the stem cell waste liquid by a centrifuge at 1800rpm/mis to remove cell flocculation membranes and salt stem cell fragments.

Further optimization; scheme 1.6: all remaining waste liquid is aseptically collected and stored in a 4-6 refrigerator or freezer.

The method comprises the following steps: in particular to a proportional process flow for sterile and nontoxic detection of different raw materials, detection and protection requirements in the detection process are normative implemented, and attention and emergency treatment methods are provided.

Further optimization; scheme 2.2: the resin must be tested for bacteria and toxicity at a ratio of 1:10 (1 g resin, 10 g ultrapure water without CO2, mixed and taken out, then placed in a bacteria culture medium, planted in a CO2 incubator, CO 25 ℃, and cultured at 37 ℃ for 24H). Post-observation ensures sterility. According to the proportion of 1: 10: resin 1 g, CO 2-free ultrapure water 10 g, and was taken out for rapid virus detection. After the sterile and nontoxic operation is ensured, the operation environment is entered, and then various subsequent operations are carried out.

Further optimization; scheme 2.3: the aseptic operation must be done while receiving and performing the detection on the resin: (masks, goggles, gloves, protective clothing, instruments before and after 75% alcohol disinfection operation, table contact surfaces and spaces).

Further optimization; scheme 2.5: the glass bottle can is not damaged or defected, if the damage or the defection occurs or occurs in the operation process, the whole operation process can be restarted after the disinfection and the sterilization are started from the beginning, and the polluted waste liquid is required to be treated immediately and is not stored.

The method comprises the following steps: and (3) performing re-decoloring process flow, process operation method, process notes and process tracking notes on the purified waste liquid by the purified resin according to the proportion.

Further optimization; in the scheme 3.1, resin and waste liquid are subjected to 1 percent (namely, resin 1: waste liquid 100 (marked according to the content of the waste liquid)) ratio calculation in a sterile environment at the same time and then are placed in a sterile operation table.

Further, the method comprises the following steps of; scheme 3.2: the redundant resin and the waste liquid (marked according to the content of the waste liquid) are aseptically stored and are ready for later use, and the phenomenon that the resin and the waste liquid move randomly and do not move randomly is avoided during the period, so that the product is prevented from being infected by dust particles and bacteria in the environment.

Further optimization; scheme 3.3: the same sterile glass bottle is taken, and the volume is calculated according to the scale or the volume and then added according to the proportion of 1 percent (namely, the resin 1 and the waste liquid 100 (marked according to the content of the waste liquid)) into the sterile glass bottle.

Further optimization; scheme 3.4: and (3) transferring the matched waste liquid bottles and cans filled with the waste liquid (marked according to the content of the waste liquid) to a 4-8-degree refrigerator for storage or a sterile operation device for standing and storage. Visual changes were observed in the morning and evening every day starting a week; the color starts to change and thus a recording is made. The next day of the second week, the waste started to become reddish-mixed, and was visually observed once a day and recorded. Starting to fade the color on the second day of the third week, namely, visually observing once on two days and recording; the colour started to sparkle over the weekend. The colour was clear at the beginning of the fourth week, and a record of one week was made during this time with the colour removal precipitation.

Further optimization; scheme 3.5: the complete transparent layer and the impurity layer of the waste liquid with different contents (marked according to the content of the waste liquid) can be seen to form obvious contrast in the fourth period, the record is also made at the moment, and the picture is compared and stored at the necessary period.

Further optimization 3.6: the fibers were clear during this week of visual inspection, and after filtration, they were left for 48 hours and were clearly visible in a microscope as tiny floc fibers. For these fibers there are respectively: protein fibers, factor fibers, cytoplasmic fibers.

And 3.2, performing re-decolorization, process operation, process notes and process tracking notes on the purified waste liquid by using the purified activated carbon according to the proportion according to the process flow, proportion, operation specification and notes of activated carbon detection. Aims to ensure that various factors and proteins secreted by stem cells in the purified essence are substantially protected from being damaged by activity, 0.1-0.3% of food and drug grade preservative is added, and the essence is stored at a low temperature of 4-6 ℃. Is suitable for medical, American and medical products.

Further, the method comprises the following steps of; scheme 3.7: and adding activated carbon (90 percent to 96 percent to 98 percent) in the original proportion into the stock solution product filtered from the stem cell waste liquid, wherein the activated carbon 1 and the raw material stock solution product are 100 percent. The activated carbon of the method has the corresponding grade qualification, and the method can be operated, and the activated carbon detection comprises the following steps: according to the proportion of 1:10 (1 g of activated carbon, 10 g of ultrapure water without CO2, mixed and taken out, then placed in a bacterial culture medium, implanted in a CO2 incubator, CO 25 ℃ and subjected to bacterial culture at the temperature of 37 ℃ for 24H). Post-observation ensures sterility. According to the proportion of 1: 10: 1 g of activated carbon and 10 g of ultrapure water without CO2, and taking out for rapid virus detection test. After the sterile and nontoxic operation is ensured, the operation environment is entered, and then various subsequent operations are carried out.

Further optimization; scheme 3.8: the initial week was visually observed to be a dark brown color with a slight cloudiness in the middle, and the initial week was recorded once a day, morning and night. The next day of the second week, the waste started to become reddish-mixed, and was visually observed once a day and recorded. Starting to fade the color on the second day of the third week, namely, visually observing once on two days and recording; the colour started to sparkle over the weekend. The color was clear at the beginning of the fourth week, and a record of one week was made during this time with the decolorizing precipitation.

Further optimization; scheme 3.9: the sample after standing is obviously glittering and translucent under a microscope, and the excessive and residual intractable salt in the sample is fully adsorbed in the activated carbon and wrapped in the impurities on the ground.

Further optimization; in the scheme 3.10, under the normal condition that all factors and proteins of the purified essence are protected, the purified essence is filtered by a precise filter screen, then is precipitated for 2 weeks, and is collected with ultrapure essence, the precipitate is removed, 0.1-0.3% of food-grade and pharmaceutical-grade preservative is added, and the mixture is placed at a low temperature of 4-6 ℃ for preservation. The product is suitable for medical and American products and medical grade products, and is added and blended at the low temperature of 20-30 ℃ according to the proportion.

Further optimization; scheme 3.11: all operating specifications, operating procedures must be performed aseptically and strictly: the mask, goggles, gloves, protective clothing, instrument and table contact surfaces and spaces before and after 75% alcohol disinfection operation ensure artificial fungus implantation. When the articles are scattered on the ground in the operation process, the articles do not need to be picked up by stretching hands, and after the operation is finished, the articles are collected in the waste bag and are cleaned out.

Further optimization; scheme 3.12: the PH of the raw material stock solution can be adjusted to the compound required degree in the buffer solution; it is generally recommended that between PH6-7.5, more next-step executions can be performed to meet each need.

The invention relates to a stem cell waste liquid precipitation decoloring method and application thereof, wherein the method comprises the following steps: the stem cell experimental medical waste liquid is subjected to a series of decoloration, purification and purification to obtain the raw material level essence liquid internal nutrition: the activity, stability and safe landing of protein, factors and dismutase all meet the requirements of activity repair, activation and regeneration of skin cells and epidermal cells; the excess salt, floccules and impurities in the stem cell waste liquid are effectively adsorbed and post-treated in a decoloring, purifying and purifying sequence; the invention comprises the following steps: a method for decolorizing stem cell waste liquid by precipitation and application thereof; the raw material level essence is easy to add and is comprehensively and easily dissolved in oil and water.

Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.