阅读说明：本技术 一种免疫细胞冻存装置及其使用方法 (Immune cell cryopreservation device and using method thereof ) 是由 袁曾津 傅小聪 于 2021-07-07 设计创作，主要内容包括：本发明公开了一种免疫细胞冻存装置,其技术方案要点是：包括盒体,还包括：盖体,所述盖体的一侧与所述盒体铰接；锁合机构,所述盖体的另一侧通过所述锁合机构与所述盒体固定连接；放置板,所述放置板设置在所述盒体内；放置腔,所述放置腔设置在所述放置板内,且所述放置腔的底部设置在所述盒体内；冻存管,所述冻存管插接在所述放置腔内；密封盖,所述密封盖螺纹连接在所述冻存管的顶部；本发明还公开了一种免疫细胞冻存装置的使用方法。本发明通过冻存管对免疫细胞进行存放,然后将冻存管放入放置腔内,并将放置板放入盒体内部,最后利用锁合机构将盒体和盖体固定,从而增加了免疫细胞存放时的安全性,有利于免疫细胞的保存和运输。(The invention discloses an immune cell cryopreservation device, which has the technical scheme that: including the box body, still include: the cover body, one side of the said cover body is hinged with said box body; the other side of the cover body is fixedly connected with the box body through the locking mechanism; the placing plate is arranged in the box body; the placing cavity is arranged in the placing plate, and the bottom of the placing cavity is arranged in the box body; the freezing tube is inserted into the placing cavity; the sealing cover is in threaded connection with the top of the freezing storage pipe; the invention also discloses a using method of the immune cell cryopreservation device. According to the invention, the immune cells are stored through the freezing storage tube, then the freezing storage tube is placed in the placing cavity, the placing plate is placed in the box body, and finally the box body and the cover body are fixed by using the locking mechanism, so that the safety of the immune cells during storage is increased, and the immune cells are favorably stored and transported.)

1. The utility model provides an immune cell cryopreserving device, includes box body (1), its characterized in that still includes:

the box body (1) is hinged with one side of the cover body (2);

the other side of the cover body (2) is fixedly connected with the box body (1) through the locking mechanism (3);

a placement plate (4), the placement plate (4) being disposed within the box body (1);

the placing cavity (5) is arranged in the placing plate (4), and the bottom of the placing cavity (5) is arranged in the box body (1);

the freezing tube (6), the freezing tube (6) is inserted in the placing cavity (5);

the sealing cover (7) is in threaded connection with the top of the freezing storage pipe (6), and the lower surface of the cover body (2) is in contact with the upper surface of the sealing cover (7).

2. The device of claim 1, wherein the frozen immune cell storage device comprises: the locking mechanism (3) comprises two groups of fixing blocks (31), and the two groups of fixing blocks (31) are fixedly arranged on the surface of the cover body (2);

the locking plate (32) is hinged between the two groups of fixing blocks (31);

a through groove (33), wherein the through groove (33) is arranged on the locking plate (32);

the locking lug (34) is fixedly arranged on the surface of the box body (1), and the locking lug (34) penetrates through the through groove (33);

copper lock (35), locking plate (32) pass through copper lock (35) with lock ear (34) fixed connection.

3. The device of claim 1, wherein the frozen immune cell storage device comprises: place chamber (5) with it all is provided with sixteen groups to freeze and deposits pipe (6), sixteen groups place chamber (5) and sixteen groups it all is 4x4 matrix arrangement to freeze and deposit pipe (6).

4. The device of claim 1, wherein the frozen immune cell storage device comprises: the clamping blocks (8) are symmetrically arranged on two sides of the placing plate (4), clamping grooves (9) are symmetrically formed in two sides of the box body (1), and the clamping grooves (9) are formed in the clamping grooves (9).

5. The device of claim 1, wherein the frozen immune cell storage device comprises: the edge of the placing plate (4) is hermetically connected with the inner wall of the box body (1).

6. The device of claim 1, wherein the frozen immune cell storage device comprises: the box body (1) is square, the bottom of the box body (1) is of a transparent structure, and scale marks are arranged on the outer surface of the box body (1).

7. A use method of an immune cell cryopreservation device is characterized in that: the method comprises the following steps:

s1, filling isopropanol into the box body (1) for later use according to scale marks on the box body (1);

s2, placing the placing plate (4) on the box body (1), and inserting the bottom of the placing cavity (5) into isopropanol;

s3, preparing immune cell cryopreservation suspension;

s4, filling the immune cell cryopreservation suspension into a cryopreservation tube (6), and covering a sealing cover (7);

s5, putting the freezing tube (6) into the placing cavity (5), and covering the cover body (2);

s6, fixing the cover body (2) and the box body (1) by using the locking mechanism (3);

s7, the cover body (2) and the box body (1) are integrally placed into a low-temperature refrigerator at minus 80 ℃ for storage.

8. The method of claim 7, wherein the frozen immune cell storage device comprises: the preparation method of the isopropanol comprises the following steps:

s101, in the presence of a strong acid solid catalyst, directly hydrating propylene and water, separating a crude alcohol aqueous solution from an obtained reaction product, and refining the aqueous solution;

s102, continuously adding propylene, water and saturated hydrocarbon into a reactor, wherein the solid catalyst is filled or suspended in the reactor, and each mole of propylene to be reacted needs 1-3 moles of water;

s103, carrying out hydration reaction while keeping the temperature and the pressure in the reactor equal to the critical temperature and the critical pressure of the propylene and the saturated hydrocarbon;

s104, recycling the obtained reaction product to an inlet of the reactor;

s105, discharging all vapor phases from reaction products, unreacted raw materials and saturated hydrocarbon from the reactor, liquefying isopropanol and water by reducing pressure and cooling, separating vapor components, and refining crude isopropanol contained in the obtained liquefied phase to obtain refined isopropanol.

9. The method of claim 8, wherein the cryopreservation device comprises: the preparation method of the isopropanol comprises the following steps:

s201, placing strong acid cation exchange resin in a low-carbon alcohol solvent, soaking for 7-28 h at 40-160 ℃, then introducing inert gas in three stages, wherein the conditions of introducing the inert gas in the first stage are that the pressure is 0.4-0.7 MPa, the temperature is 45-65 ℃, and the flow rate of the inert gas is 1m³/h-3m³H; the conditions of the second stage of introducing inert gas are that the pressure is 0.7MPa to 1.2MPa, the temperature is 65 ℃ to 85 ℃, and the flow rate of the inert gasIs 3m³/h-6m³H; the third stage is fed with inert gas under the conditions of pressure of 1.2MPa-1.6MPa, temperature of 85-110 deg.C and inert gas flow rate of 6m³/h-9m³Preparing a modified strong acid cation exchange resin catalyst;

s202, hydrolyzing isopropyl acetate and water under the action of the modified strong acid cation exchange resin catalyst of S201 to generate a mixture of isopropanol and acetic acid, separating to obtain isopropanol, and concentrating the acetic acid for recycling.

10. The method of claim 9, wherein the cryopreservation device comprises: the strong acid cation exchange resin is D005 type strong acid cation exchange resin; the low-carbon alcohol solvent is normal alcohol of C1-C7.

Technical Field

The invention relates to the technical field of cell storage, in particular to an immune cell cryopreservation device and a using method thereof.

Background

Immune cells refer to cells involved in or associated with an immune response. Including lymphocytes, dendritic cells, monocytes/macrophages, granulocytes, mast cells, and the like. Immune cells can be classified into various types, and various immune cells play an important role in the human body. Immune cells (immuno cells) are commonly referred to as leukocytes, and include innate lymphocytes, various phagocytic cells, and the like, and lymphocytes that recognize antigens and generate specific immune responses, and the like.

Referring to the prior Chinese patent with the publication number of CN112674078A, the ultrasonic cryopreservation device for hepatocyte ice-planting and the cryopreservation method thereof are disclosed, and comprise an ultrasonic container, a temperature detection system, an ultrasonic induced nucleation system and a cooling system; the ultrasonic liver cell ice-planting cryopreservation method comprises the following steps: culturing hepatic cells, determining the upper threshold of the ice-planting temperature, selecting raw materials of temperature-controlled phase-change liquid, and carrying out ultrasonic ice-planting on the hepatic cells for freezing storage.

The ultrasonic hepatocyte ice planting cryopreservation device and the cryopreservation method thereof obviously improve the cryopreservation effect of hepatocytes, the consumption of cryoprotectant is small, the ultrasonic ice planting and the cryopreservation protectant have a synergistic effect, the recovery survival rate of the cryopreserved hepatocytes is higher than 90%, and the cryopreservation cryoprotectant has no significant difference with a fresh group, so that an unexpected technical effect is achieved. However, the existing immune cell cryopreservation device and the use method thereof still have some disadvantages, such as: firstly, immune cells are inconvenient to store and transport; and secondly, the immune cells are not convenient to be cooled and stored slowly.

Disclosure of Invention

In view of the problems mentioned in the background art, the present invention is to provide an immune cell cryopreservation device and a method for using the same, so as to solve the problems mentioned in the background art.

The technical purpose of the invention is realized by the following technical scheme:

the utility model provides an immune cell cryopreserving device, includes the box body, still includes:

the cover body, one side of the said cover body is hinged with said box body;

the other side of the cover body is fixedly connected with the box body through the locking mechanism;

the placing plate is arranged in the box body;

the placing cavity is arranged in the placing plate, and the bottom of the placing cavity is arranged in the box body;

the freezing tube is inserted into the placing cavity;

the sealing cover is in threaded connection with the top of the cryopreservation pipe, and the lower surface of the cover body is in contact with the upper surface of the sealing cover.

Through adopting above-mentioned technical scheme, deposit immune cell through freezing the pipe of depositing, then will freeze the pipe of depositing and put into and place the intracavity to will place the board and put into inside the box body, utilize locking mechanism to fix box body and lid at last, thereby increased the security when immune cell deposits, be favorable to immune cell's save and transportation.

Preferably, the locking mechanism comprises two groups of fixed blocks, and the two groups of fixed blocks are fixedly arranged on the surface of the cover body;

the locking plate is hinged between the two groups of fixing blocks;

the through groove is formed in the locking plate;

the locking lug is fixedly arranged on the surface of the box body and penetrates through the through groove;

the copper lock, the locking plate passes through the copper lock with lock ear fixed connection.

Through adopting above-mentioned technical scheme, pass logical groove with the lock lug through rotating the locking plate, then utilize the copper lock to fix locking plate and lock lug to the convenience is fixed box body and lid, makes things convenient for the cell to preserve.

Preferably, the placing cavities and the freezing pipes are provided with sixteen groups, and the sixteen groups of the placing cavities and the sixteen groups of the freezing pipes are arranged in a 4x4 matrix.

By adopting the technical scheme, the one-time cryopreservation quantity of the immune cells is increased.

Preferably, clamping blocks are symmetrically arranged on two sides of the placing plate, clamping grooves are symmetrically formed in two sides of the box body, and the clamping grooves are formed in the clamping grooves.

Through adopting above-mentioned technical scheme, through setting up draw-in groove and fixture block, not only realize the draw-in groove spacing to the fixture block, the draw-in groove can conveniently take out the fixture block moreover to conveniently place getting of board and put.

Preferably, the edge of the placing plate is hermetically connected with the inner wall of the box body.

Through adopting above-mentioned technical scheme, increase the leakproofness, prevent the inside liquid outflow of box body.

Preferably, the box body is square, the bottom of the box body is of a transparent structure, and the outer surface of the box body is provided with scale marks.

Through adopting above-mentioned technical scheme, conveniently observe the box body is inside, and conveniently deposit the cell.

The invention also provides a using method of the immune cell cryopreservation device, which comprises the following steps:

s1, according to scale marks on a box body, filling isopropanol into the box body for later use;

s2, placing the placing plate on the box body, and inserting the bottom of the placing cavity into the isopropanol;

s3, preparing immune cell cryopreservation suspension;

s4, filling the immune cell cryopreservation suspension into a cryopreservation tube, and covering a sealing cover;

s5, placing the freezing tube into the placing cavity, and covering the cover body;

s6, fixing the cover body and the box body by using a locking mechanism;

s7, placing the cover body and the box body into a low-temperature refrigerator at minus 80 ℃ for storage.

Through adopting above-mentioned technical scheme, through adding isopropanol in the inside of box body, can conveniently carry out the effect of slow cooling to immune cell, be favorable to improving cell survival rate simultaneously.

Preferably, the preparation method of the isopropanol comprises the following steps:

s101, in the presence of a strong acid solid catalyst, directly hydrating propylene and water, separating a crude alcohol aqueous solution from an obtained reaction product, and refining the aqueous solution;

s102, continuously adding propylene, water and saturated hydrocarbon into a reactor, wherein the solid catalyst is filled or suspended in the reactor, and each mole of propylene to be reacted needs 1-3 moles of water;

s103, carrying out hydration reaction while keeping the temperature and the pressure in the reactor equal to the critical temperature and the critical pressure of the propylene and the saturated hydrocarbon;

s104, recycling the obtained reaction product to an inlet of the reactor;

s105, discharging all vapor phases from reaction products, unreacted raw materials and saturated hydrocarbon from the reactor, liquefying isopropanol and water by reducing pressure and cooling, separating vapor components, and refining crude isopropanol contained in the obtained liquefied phase to obtain refined isopropanol.

By adopting the technical scheme, the preparation purity of the isopropanol can be increased, and the production efficiency of the isopropanol is conveniently improved.

Preferably, the preparation method of the isopropanol comprises the following steps:

s201, placing strong acid cation exchange resin in a low-carbon alcohol solvent, soaking for 7-28 h at 40-160 ℃, then introducing inert gas in three stages, wherein the conditions of introducing the inert gas in the first stage are that the pressure is 0.4-0.7 MPa, the temperature is 45-65 ℃, and the flow rate of the inert gas is 1m³/h-3m³H; the conditions of the second stage of introducing inert gas are that the pressure is 0.7MPa to 1.2MPa, the temperature is 65 ℃ to 85 ℃, and the flow rate of the inert gas is 3m³/h-6m³H; the third stage is fed with inert gas under the conditions of pressure of 1.2MPa-1.6MPa, temperature of 85-110 deg.C and inert gas flow rate of 6m³/h-9m³Preparing a modified strong acid cation exchange resin catalyst;

s202, hydrolyzing isopropyl acetate and water under the action of the modified strong acid cation exchange resin catalyst of S201 to generate a mixture of isopropanol and acetic acid, separating to obtain isopropanol, and concentrating the acetic acid for recycling.

By adopting the technical scheme, the supply of the isopropanol is convenient, and the utilization efficiency of resources is increased.

Preferably, the strong acid cation exchange resin is a D005 type strong acid cation exchange resin; the low-carbon alcohol solvent is normal alcohol of C1-C7.

In summary, the invention mainly has the following beneficial effects:

the immune cells are stored through the cryopreservation tube, then the cryopreservation tube is placed in the placing cavity, the placing plate is placed in the box body, and finally the box body and the cover body are fixed through the locking mechanism, so that the safety of the immune cells during storage is improved, and the immune cells are favorably stored and transported;

secondly, in the method, the isopropanol is added into the box body, so that the effect of slowly cooling the immune cells can be conveniently realized, and the survival rate of the cells can be improved.

Drawings

FIG. 1 is a schematic structural view of the present invention;

FIG. 2 is a structural cross-sectional view of the present invention;

FIG. 3 is a top plan view of the cartridge of FIG. 2 with the portion of the vial removed;

FIG. 4 is a schematic structural view of the locking mechanism of the present invention;

fig. 5 is a block flow diagram of the present invention.

Reference numerals: 1. a box body; 2. a cover body; 3. a locking mechanism; 31. a fixed block; 32. a locking plate; 33. a through groove; 34. locking lugs; 35. copper lock; 4. placing the plate; 5. a placement chamber; 6. freezing and storing the tube; 7. a sealing cover; 8. a clamping block; 9. a clamping groove.

Detailed Description

The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in 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.

Example 1

Referring to fig. 1-4, an immune cell cryopreservation device comprises a box body 1, and further comprises:

the cover body 2, one side of the cover body 2 is hinged with the box body 1;

the other side of the cover body 2 is fixedly connected with the box body 1 through the locking mechanism 3;

a placing plate 4, wherein the placing plate 4 is arranged in the box body 1;

the placing cavity 5 is arranged in the placing plate 4, and the bottom of the placing cavity 5 is arranged in the box body 1;

the freezing tube 6 is inserted in the placing cavity 5;

and the sealing cover 7 is in threaded connection with the top of the freezing storage pipe 6, and the lower surface of the cover body 2 is in contact with the upper surface of the sealing cover 7.

Referring to fig. 4, the locking mechanism 3 includes two fixing blocks 31, and the two fixing blocks 31 are disposed in two groups, and the two groups of fixing blocks 31 are fixedly disposed on the surface of the cover 2;

the locking plate 32 is hinged between the two groups of fixed blocks 31;

a through groove 33, wherein the through groove 33 is formed on the locking plate 32;

the locking lug 34 is fixedly arranged on the surface of the box body 1, and the locking lug 34 penetrates through the through groove 33;

the copper lock 35, the locking plate 32 is fixedly connected with the lock lug 34 through the copper lock 35; the lock plate 32 is rotated to enable the lock lug 34 to penetrate through the through groove 33, and then the copper lock 35 is used for fixing the lock plate 32 and the lock lug 34, so that the box body 1 and the cover body 2 are conveniently fixed, and the cells are conveniently stored.

Referring to fig. 3, in order to increase the number of the immune cells frozen at one time, sixteen groups of the placing cavities 5 and the freezing tubes 6 are arranged, and the sixteen groups of the placing cavities 5 and the sixteen groups of the freezing tubes 6 are arranged in a 4x4 matrix.

Referring to fig. 3, two sides of the placing plate 4 are symmetrically provided with clamping blocks 8, two sides of the box body 1 are symmetrically provided with clamping grooves 9, and the clamping grooves 9 are arranged in the clamping grooves 9; through setting up draw-in groove 9 and fixture block 8, not only realize draw-in groove 9 spacing to fixture block 8, draw-in groove 9 can conveniently take out fixture block 8 moreover to conveniently place getting of board 4 and put.

Referring to fig. 2, in order to increase the sealability, the liquid inside the case 1 is prevented from flowing out; the edge of the placing plate 4 is hermetically connected with the inner wall of the box body 1.

Referring to fig. 1, the inside of the case 1 is conveniently observed, and cells are conveniently stored; the box body 1 is square, the bottom of the box body 1 is of a transparent structure, and scale marks are arranged on the outer surface of the box body 1.

Example 2

Referring to fig. 5, a method for using an immune cell cryopreservation device comprises the following steps:

s1, filling isopropanol into the box body 1 for later use according to scale marks on the box body 1;

s2, placing the placing plate 4 on the box body 1, and inserting the bottom of the placing cavity 5 into the isopropanol;

s3, preparing immune cell cryopreservation suspension;

s4, filling the immune cell cryopreservation suspension into a cryopreservation tube 6, and covering a sealing cover 7;

s5, placing the freezing tube 6 into the placing cavity 5, and covering the cover body 2;

s6, fixing the cover body 2 and the box body 1 by using the locking mechanism 3;

s7, the cover body 2 and the box body 1 are integrally placed in a low-temperature refrigerator at minus 80 ℃ for storage.

In this embodiment, preferably, the preparation method of isopropanol includes the following steps:

s101, in the presence of a strong acid solid catalyst, directly hydrating propylene and water, separating a crude alcohol aqueous solution from an obtained reaction product, and refining the aqueous solution;

s102, continuously adding propylene, water and saturated hydrocarbon into a reactor, wherein the solid catalyst is filled or suspended in the reactor, and each mole of propylene to be reacted needs 3 moles of water;

s103, carrying out hydration reaction while keeping the temperature and the pressure in the reactor equal to the critical temperature and the critical pressure of the propylene and the saturated hydrocarbon;

s104, recycling the obtained reaction product to an inlet of the reactor;

s105, discharging all vapor phases from reaction products, unreacted raw materials and saturated hydrocarbon from the reactor, liquefying isopropanol and water by reducing pressure and cooling, separating vapor components, and refining crude isopropanol contained in the obtained liquefied phase to obtain refined isopropanol.

Example 3

The difference from example 2 is that: the preparation method of the isopropanol comprises the following steps:

s201, placing the strong acid cation exchange resin in a low-carbon alcohol solvent, soaking for 28h at 40 ℃, then introducing inert gas in three stages, wherein the conditions of introducing the inert gas in the first stage are that the pressure is 0.4MPa, the temperature is 45 ℃, and the flow rate of the inert gas is 1m³H; the second stage is fed with inert gas under the conditions of 0.7MPa pressure, 65 deg.C temperature and 3m inert gas flow rate³H; the third stage is fed with inert gas under the conditions of 1.2MPa pressure, 85 deg.C temperature and inert gas flow rate of 6m³Preparing a modified strong acid cation exchange resin catalyst;

s202, hydrolyzing isopropyl acetate and water under the action of the modified strong acid cation exchange resin catalyst of S201 to generate a mixture of isopropanol and acetic acid, separating to obtain isopropanol, and concentrating the acetic acid for recycling.

In this embodiment, preferably, the strong acid cation exchange resin is a D005 type strong acid cation exchange resin; the low-carbon alcohol solvent is normal alcohol of C1-C7.

Example 4

The difference from example 3 is that: the preparation method of the isopropanol comprises the following steps:

s201, placing the strong acid cation exchange resin in a low-carbon alcohol solvent, soaking for 7 hours at 160 ℃, then introducing inert gas in three stages, wherein the conditions of introducing the inert gas in the first stage are that the pressure is 0.7MPa, the temperature is 65 ℃, and the flow rate of the inert gas is 3m³H; the second stage is fed with inert gas under the conditions of 1.2MPa pressure, 85 deg.C temperature and 6m inert gas flow rate³H; first, theThe inert gas is introduced into the three stages under the conditions of 1.6MPa of pressure, 110 ℃ of temperature and 9m of flow speed of the inert gas³Preparing a modified strong acid cation exchange resin catalyst;

s202, hydrolyzing isopropyl acetate and water under the action of the modified strong acid cation exchange resin catalyst of S201 to generate a mixture of isopropanol and acetic acid, separating to obtain isopropanol, and concentrating the acetic acid for recycling.

In this embodiment, preferably, the strong acid cation exchange resin is a D005 type strong acid cation exchange resin; the low-carbon alcohol solvent is normal alcohol of C1-C7.

Example 5

The difference from example 3 is that: the preparation method of the isopropanol comprises the following steps:

s201, placing the strong acid cation exchange resin in a low-carbon alcohol solvent, soaking for 20h at 100 ℃, then introducing inert gas in three stages, wherein the conditions of introducing the inert gas in the first stage are that the pressure is 0.6MPa, the temperature is 55 ℃, and the flow rate of the inert gas is 2m³H; the second stage is fed with inert gas under the conditions of 0.9MPa of pressure, 75 ℃ of temperature and 5m of flow velocity of inert gas³H; the third stage is fed with inert gas under the conditions of 1.4MPa pressure, 95 deg.C temperature and 8m inert gas flow rate³Preparing a modified strong acid cation exchange resin catalyst;

s202, hydrolyzing isopropyl acetate and water under the action of the modified strong acid cation exchange resin catalyst of S201 to generate a mixture of isopropanol and acetic acid, separating to obtain isopropanol, and concentrating the acetic acid for recycling.

In this embodiment, preferably, the strong acid cation exchange resin is a D005 type strong acid cation exchange resin; the low-carbon alcohol solvent is normal alcohol of C1-C7.

The use principle and the advantages are as follows:

the immune cells are stored through the freezing tube 6, then the freezing tube 6 is placed into the placing cavity 5, the placing plate 4 is placed into the box body 1, and finally the box body 1 and the cover body 2 are fixed through the locking mechanism 3, so that the safety of the immune cells during storage is improved, and the immune cells are favorably stored and transported; according to the method, the isopropanol is added into the box body 1, so that the effect of slowly cooling the immune cells can be conveniently achieved, and the survival rate of the cells can be improved.

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.