阅读说明：本技术 一种构建小鼠同种异体皮肤移植模型的方法 (Method for constructing mouse allogeneic skin transplantation model ) 是由 于济彰 夏家红 吴杰 崔济开 许恒 陈漳 李媛 张曦 张丹 于 2021-07-29 设计创作，主要内容包括：本发明公开了一种构建小鼠同种异体皮肤移植模型的方法,包括以下步骤：S1、供体准备：处死Balb/c小鼠一只,备皮,消毒后去全部背部皮肤,刮除皮下筋膜组织及淋巴结,制备皮片,于冰生理盐水中保存；S2、受体准备：对受体小鼠进行戊巴比妥腹腔麻醉,麻醉状态满意后备皮,用脱毛膏刮除毛发,并用酒精再次消毒,剪除1cm直径圆形皮肤形成移植区；S3、皮肤移植：将皮片剪出同样大小的皮片贴于移植区；S4、固定：使用PU医用透明防水辅料固定皮片并；最后再通过医用胶布和自粘性医用绷带再次固定。通过上述设置,本方法不采用缝合的方法,并且在造模的过程中能够通过PU医用透明防水辅料观察移植状态,因此就有造模块和造模成功率高的优点。(The invention discloses a method for constructing a mouse allogeneic skin transplantation model, which comprises the following steps: s1, donor preparation: killing one Balb/c mouse, preparing skin, removing all back skin after disinfection, scraping subcutaneous fascia tissues and lymph nodes, preparing skin sheets, and storing in ice physiological saline; s2, receptor preparation: carrying out pentobarbital abdominal anesthesia on a receptor mouse, preparing skin after the anesthesia state is satisfied, scraping hair by depilatory cream, disinfecting by alcohol again, and cutting out 1 cm-diameter circular skin to form a transplantation area; s3, skin grafting: cutting the skin into pieces with the same size and sticking the pieces to the transplanting area; s4, fixing: fixing the leather sheets by using PU medical transparent waterproof auxiliary materials; and finally, fixing again through the medical adhesive plaster and the self-adhesive medical bandage. Through the arrangement, the method does not adopt a sewing method, and the transplanting state can be observed through the PU medical transparent waterproof auxiliary material in the molding process, so that the method has the advantages of high molding and molding success rate.)

1. A method for constructing a mouse allogeneic skin transplantation model is characterized by comprising the following steps: the method comprises the following steps:

s1, donor preparation: killing one Balb/c mouse, preparing skin, removing all back skin after disinfection, scraping subcutaneous fascia tissues and lymph nodes, preparing a skin sheet (1), and storing in ice physiological saline;

s2, receptor preparation: carrying out pentobarbital abdominal anesthesia on a receptor mouse, preparing skin after the anesthesia state is satisfied, scraping hair by depilatory cream, disinfecting by alcohol again, and cutting off round skin to form a transplantation area (2);

s3, skin grafting: cutting the skin sheet (1) into the same size as the transplanting area (2), and sticking the skin sheet to the transplanting area (2);

s4, fixing: fixing the skin piece (1) by using a PU medical transparent waterproof auxiliary material and finely adjusting the position of the skin piece (1) under the condition of direct vision of naked eyes so that the skin piece (1) and the transplanting area (2) are in the best fitting state; fixing the PU medical transparent waterproof auxiliary material by using a medical adhesive tape (6), wherein the fixed position is deviated from the transplanting area (2); the compression fixation is carried out again using a self-adhesive medical bandage (7) without affecting the respiration of the mice.

2. The method of claim 1, wherein the method comprises the steps of: the diameter of the transplanting area (2) is 1cm-3 cm.

3. The method of claim 1, wherein the method comprises the steps of: the PU medical transparent waterproof auxiliary material is cut into a fixed area (3), and the fixed area (3) is round or is deformed regularly.

4. The method of claim 3, wherein the method comprises the steps of: the center of the fixed area (3) is cut with a center hole (4) with the diameter smaller than that of the transplanting area (2).

5. The method of claim 4, wherein the method comprises the steps of: the outer side of the fixed area (3) is uniformly provided with a fixed belt (5) formed by shearing.

6. The method of claim 5, wherein the method comprises the steps of: the medical adhesive plaster (6) is fixedly arranged on the fixing band (5).

Technical Field

The invention relates to the field of medical animal models, in particular to a method for constructing a mouse allogeneic skin transplantation model.

Background

In 1943, Medawar constructed a mouse allogeneic skin graft model for the first time, and revealed the basic rule that autologous skin graft would not be rejected, whereas allogeneic skin graft would be rejected. The establishment of the skin transplantation model lays a foundation for the development of modern transplantation and has important significance. In recent years, skin-containing grafts such as clinical hand graft, face graft, abdominal wall graft, penis graft, etc. have been rapidly spread, but patients still need higher doses of immunosuppressive agents due to high immunogenicity of skin. The mouse skin graft model has been established for nearly 70 years, but the mechanism of skin graft rejection is still not completely understood, and therefore, a suitable mouse allogeneic skin graft model is crucial for further studies on the mechanism of skin rejection.

For example, the invention patent application with the application number of 2015104871735 discloses a mouse vascularization skin transplantation model and a construction method thereof, wherein an abdominal skin flap with a mouse femoral artery and vein as a pedicle is transferred to the neck of another mouse, the femoral artery and the common carotid artery are connected by using a vascular sleeve (cuff) technology, and the femoral vein and the external jugular vein are connected; the invention aims to provide a mouse vascularized skin transplantation model with simple and convenient operation, good repeatability and high success rate and a construction method thereof, which can be used for researching the clinical vascularized skin transplantation rejection mechanism and the animal model for inducing the immune tolerance mechanism.

However, the above construction method has not been widely used due to the high operation difficulty. Currently, there are two general approaches to the construction of mouse models: 1. the pressure dressing method without sewing has the defects of low success rate, about 50 percent of success rate of a new hand and about 80 percent of success rate after proficiency. 2. The sewing and binding method has the defects of slow molding time and certain technical requirements on experimenters for sewing.

In view of the above problems, the present application provides a method for constructing a mouse allogeneic skin graft model.

Disclosure of Invention

The invention aims to provide a method for constructing a mouse allogeneic skin transplantation model, which does not adopt a sewing method and can observe the transplantation state through PU medical transparent waterproof auxiliary materials in the molding process, thereby having the advantages of high module construction and molding success rate.

In order to achieve the purpose, the invention provides the following technical scheme: a method for constructing a mouse allogeneic skin transplantation model comprises the following steps:

s1, donor preparation: killing one Balb/c mouse, preparing skin, removing all back skin after disinfection, scraping subcutaneous fascia tissues and lymph nodes, preparing skin sheets, and storing in ice physiological saline;

s2, receptor preparation: carrying out pentobarbital abdominal anesthesia on a receptor mouse, preparing skin after the anesthesia state is satisfied, scraping hair by depilatory cream, disinfecting by alcohol again, and cutting off round skin to form a transplantation area;

s3, skin grafting: cutting the skin into pieces with the same size as the transplanting area, and sticking the pieces to the transplanting area;

s4, fixing: fixing the skin sheet by using the PU medical transparent waterproof auxiliary material and finely adjusting the position of the skin sheet under the condition of direct vision of naked eyes so that the skin sheet and the transplanting area are in the best fitting state; fixing the PU medical transparent waterproof auxiliary material by using a medical adhesive tape, wherein the fixed position is deviated from the transplanting area; the compression fixation is carried out again by using a self-adhesive medical bandage under the condition of not influencing the breathing of the mouse.

Preferably, the diameter of the graft region is 1cm to 3 cm.

Preferably, the PU medical transparent waterproof auxiliary material is cut into a fixed area, and the fixed area is circular or is deformed regularly.

Preferably, the central cut of the fixation region has a central hole with a diameter smaller than that of the graft region.

Preferably, the outer side of the fixing area is uniformly provided with fixing belts formed by shearing.

Preferably, the medical adhesive plaster is fixedly arranged on the fixing belt.

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

(1) the method does not need to sew, so the method has low requirements on sewing technology, can avoid the case of molding failure caused by sewing problems, and is suitable for large-area popularization;

(2) the PU medical transparent waterproof auxiliary material is adopted to enable the transplanting area and the skin sheet to be in a visible state in the whole molding process, so that the transplanting area and the skin sheet can be in the best position when the molding starts, and problems can be found in time and adjusted in the molding process;

(3) the central hole is arranged, so that the leather sheet can be in contact with air, a certain breathing effect is generated, and the mold making success rate of the model is improved.

Drawings

FIG. 1 is a schematic view showing a connection structure of a fastening tape and a fastening region according to one embodiment;

FIG. 2 is a graph comparing survival rates in example two;

FIG. 3 is a comparison of the operation times in example two;

FIG. 4 is a graph comparing survival times in example two;

FIG. 5 is a view showing a connection structure of a fastening tape and a fastening region in accordance with a third embodiment;

FIG. 6 is a view showing a connection structure of a fastening tape and a fastening region in accordance with the fourth embodiment.

Description of the drawings: 1. a skin sheet; 2. a transplanting area; 3. a fixed zone; 4. a central bore; 5. fixing belts; 6. a medical adhesive plaster; 7. self-adhesive medical bandages.

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.

Referring to fig. 1-6, the following are examples provided by the present invention:

the first embodiment is as follows: a mouse allogeneic skin graft model constructed by the following method comprising the steps of:

s1, donor preparation: killing one Balb/c mouse, preparing skin, removing all back skin after disinfection, scraping subcutaneous fascia tissues and lymph nodes, preparing a skin sheet 1, and storing in ice physiological saline;

s2, receptor preparation: carrying out pentobarbital abdominal anesthesia on a receptor mouse, preparing skin after the anesthesia state is satisfied, scraping hair by depilatory cream, disinfecting by alcohol again, and cutting off round skin to form a transplantation area 2; the diameter of the graft region 2 is typically 1cm to 3cm, in this example 1 cm.

S3, skin grafting: cutting the skin 1 into pieces with the same size as the transplanting area 2, and sticking the pieces to the transplanting area 2;

s4, fixing:

s4.1, cutting the PU medical transparent waterproof auxiliary material into a circular or positive multi-deformation fixing area 3, wherein the fixing area 3 is circular in the embodiment, as shown in figure 1;

s4.2, the central shearing diameter of the fixing area 3 is smaller than the central hole 4 of the transplanting area 2;

s4.3, uniformly arranging the cut fixing belts 5 on the outer side of the fixing area 3, wherein the number of the fixing belts 5 is 8 in the embodiment;

s4.4, keeping the center hole 4 coincident with the center of the skin piece 1, connecting the fixing area 3 with the skin piece 1, and finely adjusting the position of the skin piece 1 under the condition of direct vision of naked eyes to enable the skin piece 1 and the transplanting area 2 to be in the best fitting state;

and S4.5, fixing the fixing belt 5 for the second time by using a medical adhesive tape 6.

And S4.6, under the condition that the breathing of the mouse is not influenced, the self-adhesive medical bandage 7 is used for pressurizing and fixing again.

In the embodiment one, cut through the medical transparent waterproof auxiliary material to PU, and then set up fixed area 3 into circularly, and set up centre bore 4 at 3 centers of circular shape fixed area, through this kind of setting, can make the medical transparent waterproof auxiliary material of annular PU play fixed effect to the junction that skin was transplanted, can keep making skin 1 and donor skin and air contact as far as possible simultaneously again, and then can keep the autonomic breathing of skin 1 and donor skin, be favorable to successfully accomplishing skin transplantation work.

Example two, mouse allograft skin graft model, the method of example one is referred to and 20 mice are molded at the same time, and the survival rate of the molding, the operation time and the success rate of the molding are counted (the comparison is only for the suture method, and the data of the pressurization method are lower than the suture method, so the details are not described herein), as shown in fig. 2, fig. 3 and fig. 4.

By analyzing the contents of fig. 2, fig. 3 and fig. 4, it can be seen that the method adopted in the present application has the advantages of high survival rate, short operation time and molding success rate, so that the molding cost can be reduced, and the method is suitable for large-scale popularization and is beneficial to deep research on skin transplantation.

Example three: a mouse allogeneic skin graft model constructed by the following method comprising the steps of:

s1, donor preparation: killing one Balb/c mouse, preparing skin, removing all back skin after disinfection, scraping subcutaneous fascia tissues and lymph nodes, preparing a skin sheet 1, and storing in ice physiological saline;

s2, receptor preparation: carrying out pentobarbital abdominal anesthesia on a receptor mouse, preparing skin after the anesthesia state is satisfied, scraping hair by depilatory cream, disinfecting by alcohol again, and cutting off round skin to form a transplantation area 2; the diameter of the graft region 2 is typically 1cm to 3cm, in this example 1 cm.

S3, skin grafting: cutting the skin 1 into pieces with the same size as the transplanting area 2, and sticking the pieces to the transplanting area 2;

s4, fixing:

s4.1, cutting the PU medical transparent waterproof auxiliary material into a round or positive multi-deformation fixing area 3, wherein the fixing area 3 is square in the embodiment, as shown in figure 5;

s4.2, the central shearing diameter of the fixing area 3 is smaller than the central hole 4 of the transplanting area 2;

s4.3, uniformly arranging the cut fixing belts 5 on the outer side of the fixing area 3, wherein the number of the fixing belts 5 is 4 in the embodiment;

s4.4, keeping the center hole 4 coincident with the center of the skin piece 1, connecting the fixing area 3 with the skin piece 1, and finely adjusting the position of the skin piece 1 under the condition of direct vision of naked eyes to enable the skin piece 1 and the transplanting area 2 to be in the best fitting state;

and S4.5, fixing the fixing belt 5 for the second time by using a medical adhesive tape 6.

And S4.6, under the condition that the breathing of the mouse is not influenced, the self-adhesive medical bandage 7 is used for pressurizing and fixing again.

In the third embodiment, the fixing area 3 is set to be square by shearing the PU medical transparent waterproof auxiliary material, and the center of the fixing area 3 of the square is provided with the central hole 4. This solution is complementary to the circular fixation area in the first embodiment. Through the setting, can make the medical transparent waterproof auxiliary material of annular PU play fixed effect to the junction that skin transplanted, the shearing degree of difficulty of the fixed area 3 of square is low simultaneously, consequently completion shearing that can be quick is favorable to improving the molding efficiency of transplanting the model. Example four: a mouse allogeneic skin graft model constructed by the following method comprising the steps of:

s1, donor preparation: killing one Balb/c mouse, preparing skin, removing all back skin after disinfection, scraping subcutaneous fascia tissues and lymph nodes, preparing a skin sheet 1, and storing in ice physiological saline;

s2, receptor preparation: carrying out pentobarbital abdominal anesthesia on a receptor mouse, preparing skin after the anesthesia state is satisfied, scraping hair by depilatory cream, disinfecting by alcohol again, and cutting off round skin to form a transplantation area 2; the diameter of the graft region 2 is typically 1cm to 3cm, in this example 1 cm.

S3, skin grafting: cutting the skin 1 into pieces with the same size as the transplanting area 2, and sticking the pieces to the transplanting area 2;

s4, fixing:

s4.1, cutting the PU medical transparent waterproof auxiliary material into a circular or positive multi-deformation fixing area 3, wherein the fixing area 3 is a regular hexagon in the embodiment, as shown in figure 6;

s4.2, the central shearing diameter of the fixing area 3 is smaller than the central hole 4 of the transplanting area 2;

s4.3, uniformly arranging the cut fixing belts 5 on the outer side of the fixing area 3, wherein the number of the fixing belts 5 is 6 in the embodiment;

s4.4, keeping the center hole 4 coincident with the center of the skin piece 1, connecting the fixing area 3 with the skin piece 1, and finely adjusting the position of the skin piece 1 under the condition of direct vision of naked eyes to enable the skin piece 1 and the transplanting area 2 to be in the best fitting state;

and S4.5, fixing the fixing belt 5 for the second time by using a medical adhesive tape 6.

And S4.6, under the condition that the breathing of the mouse is not influenced, the self-adhesive medical bandage 7 is used for pressurizing and fixing again.

In the fourth embodiment, the PU medical transparent waterproof auxiliary material is cut, so that the fixing area 3 is arranged to be a regular hexagon, and the center hole 4 is arranged at the center of the fixing area 3 of the regular hexagon. This solution is complementary to the circular and square anchoring zones 3 of the first and second embodiments. Aiming at explaining, according to specific requirements and the number of the fixing belts 6, the shape of the fixing area 3 can be flexibly selected, so that the relative fixation of the skin sheet 1 and the transplanting area 2 is enhanced as much as possible, and the mold making success rate is favorably improved.

In conclusion, the beneficial effects of the invention are as follows: (1) the method does not need to sew, so the method has low requirements on sewing technology, can avoid the case of molding failure caused by sewing problems, and is suitable for large-area popularization; (2) the PU medical transparent waterproof auxiliary material is adopted to ensure that the transplanting area 2 and the skin sheet 1 can be in a visible state in the whole molding process, so that the positions of the transplanting area and the skin sheet can be optimal when the molding is started, and problems can be found in time and adjusted in the molding process; (3) the central hole 4 is arranged, so that the leather sheet 1 can be contacted with air, a certain breathing effect is generated, and the mold making success rate of the model is improved.

It will be evident to those skilled in the art that the invention is not limited to the details of the foregoing illustrative embodiments, and that the present invention may be embodied in other specific forms without departing from the spirit or essential attributes thereof. The present embodiments are therefore to be considered in all respects as illustrative and not restrictive, the scope of the invention being indicated by the appended claims rather than by the foregoing description, and all changes which come within the meaning and range of equivalency of the claims are therefore intended to be embraced therein. Any reference sign in a claim should not be construed as limiting the claim concerned.