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A kind of stem cell skin adhesive sheet and its preparation method and application

An adhesive sheet and stem cell technology, which is applied in the field of stem cell skin adhesive sheet and its preparation, can solve the problems of inability to repair skin function, fail to realize skin function repair, cumbersome and complicated preparation process, etc., so as to improve skin repair effect and reduce skin trauma quickly. , the effect of immune rejection is not easy

Active Publication Date: 2020-08-04
SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, clinically used wound dressings are roughly divided into three categories: natural materials, animal tissue derivative products, and synthetic polymer materials. However, the inventors found that the above-mentioned wound dressings generally have different sources of insufficient sources or are prone to autoimmune rejection. Response and other shortcomings, but also can not achieve functional repair of the skin (such as no scar repair)
At the same time, in recent years, there have been more and more studies on tissue-engineered artificial skin. However, from the current research, it also faces the problem of immune rejection when using allogeneic skin, and more importantly, it cannot solve the function of the skin. Fix the problem, while the preparation process is cumbersome and complicated, and the preparation cost is high

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  • A kind of stem cell skin adhesive sheet and its preparation method and application
  • A kind of stem cell skin adhesive sheet and its preparation method and application

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preparation example Construction

[0032] In view of this, in a specific embodiment of the present invention, a kind of preparation method of stem cell skin adhesive sheet is provided, and described preparation method comprises:

[0033] S1. Separating and obtaining skin stem cells from skin tissue blocks;

[0034] S2, culture and passage expansion of skin stem cells;

[0035] S3. Lay the skin stem cells obtained through subculture and expansion on the cell membrane and incubate to obtain a cell paste sheet.

[0036] In yet another specific embodiment of the present invention, the specific method of step S1 is:

[0037] Take a piece of fresh skin tissue, soak it with 75% ethanol and 3% double antibody in PBS in turn; then chop the tissue, add digestive enzyme mixture to incubate and enzymolyze to obtain enzymatic hydrolysis solution I, and then add 0.25% trypsin to continue incubation Enzymolysis solution II was obtained, and then DNase I was added to continue incubation to obtain solution III; after the dige...

Embodiment 1

[0056] The specific steps of skin stem cell paste preparation:

[0057] 1. Isolation of skin stem cells: Take fresh skin tissue pieces weighing more than 0.1 g, treat them with 75% ethanol for 30 seconds, soak in PBS containing 3% double antibody for 3 minutes, and wash twice. Then mince the tissue with a scalpel blade, put it into a sterile EP tube, add 15ml of lyase mixture per gram of tissue (containing 4.0mg / L of lyase (dispase) and collagenase (collagenase), 37 ℃ water bath After incubation for 1 hour, mix well, then add 0.25% trypsin at a volume ratio of 1:3 and continue to incubate for 30 minutes, then add DNase I at a volume ratio of 1:1000 and continue to incubate for 5 minutes. Add an equal volume of stop solution (digestive enzyme inhibitor ) to terminate the digestion, gently pipette to obtain a single cell suspension, and filter the cell suspension with a 100 micron pore filter, take the filtered cells, resuspend and wash once with DMEM with serum, and then resusp...

Embodiment 2

[0060] Example 2 Transplantation test in vivo (nude mice) of skin sticking sheet

[0061] In order to verify that the prepared cell paste sheet has functional living skin stem cells, the prepared cell paste sheet was transplanted into nude mice with skin wounds to detect whether the wounds could heal and form complete skin.

[0062] (1) Take 8-week-old nude mice (nude / nude), after anesthesia ( figure 2 A), disinfect the skin on the back of the nude mouse, and use surgical scissors to cut off a circular area slightly smaller than the diaphragm to create a full-thickness skin wound;

[0063] (2) The prepared cell paste sheet was completely covered on the skin wound of nude mice, covered with Vaseline gauze, and bandaged (no suture needed) ( figure 2 B);

[0064] (3) The dressing tape can be removed after 10 days, and the wound has completely healed;

[0065] (4) After 1 month, raised human skin with pigment (higher than that of mouse skin) can be seen on the wound. And the...

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Abstract

The invention provides a stem cell skin patch and a preparation method and application thereof. The stem cell skin patch can be obtained by in vitro culture by using an autologous normal skin tissue (very small amount) for isolating skin stem cells (including epidermal stem cells or dermal stem cells) or other tissues to separate mesenchymal stem cells including adipose stem cells to reduce the immunological rejection caused by allogeneic skin matrix, the antigenicity of stem cell adhesion is reduced, skin repair efficiency is improved, and the potential to regenerate functional skin is achieved; and meanwhile, the preparation method is simple, complicated and expensive equipment is not required, and good practical application value is achieved.

Description

technical field [0001] The invention belongs to the technical field of biological tissue engineering, and in particular relates to a stem cell skin adhesive sheet and its preparation method and application. Background technique [0002] The information disclosed in this background section is only intended to increase the understanding of the general background of the present invention, and is not necessarily taken as an acknowledgment or any form of suggestion that the information constitutes the prior art already known to those skilled in the art. [0003] The skin is the barrier between the human body and the external environment. It not only plays the roles of protection, secretion, metabolism and sensation, but also participates in the immune response and maintains the stability of the internal environment of the body. As the most exposed skin of the human body, various severe skin ulceration diseases, burns, scalds, etc. are common injuries in daily life. After the ski...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/074A61L27/38A61L27/50A61L27/60
CPCA61L27/3834A61L27/3895A61L27/50A61L27/60C12N5/0625C12N2509/00
Inventor 吴训伟白福响冷雪
Owner SHANDONG UNIV