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A kind of culture method and culture medium of autologous tissue engineered epidermis

A tissue engineering and culture method technology, applied in the field of cell culture, can solve the problems of creating wounds, increasing damage, and high cost, and achieving the effects of promoting cell maturation and differentiation, accelerating peeling speed, and reducing damage

Active Publication Date: 2021-02-26
SHANGHAI BIOMED UNION BIOTECHNOLOGY CO LTD
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  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In 1975, Rheinwald and Green invented the method of culturing mouse embryonic fibroblast cells (3T3) as trophoblast cells. In order to solve the safety problems caused by animal-derived cell lines, many people also used autologous fibroblasts as trophoblast cells. Cultivate; This method mainly has following four defects: a) the safety problem of animal source cytotrophoblast; It is necessary to take two skins from the patient to increase the damage; d) The fibroblasts of different people have different activities and functions, and the stability of the trophoblast layer is different, and the quality of the cultured epidermal cells is difficult to guarantee
However, there are still risks in this method: a) The basal layer is easily damaged after being digested with DISPASE enzyme. Under the condition of serum-free medium, the cultured epidermal cells will not be divided into epidermal structures, and the epidermal cells need to be seeded on the scaffold material to complete the tissue Construction of engineered epidermis: b) epidermal cells are difficult to divide evenly on the scaffold material, and cannot have a complete epidermal structure and function; c) scaffold materials are mainly chemically synthesized, which is costly and increases clinical risks

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  • A kind of culture method and culture medium of autologous tissue engineered epidermis
  • A kind of culture method and culture medium of autologous tissue engineered epidermis
  • A kind of culture method and culture medium of autologous tissue engineered epidermis

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Embodiment

[0035] A kind of culture method of autologous tissue engineered epidermis of the present invention comprises the following steps:

[0036] 1. Pretreatment of epidermal tissue 1: Take normal epidermal skin 2cm*2cm, wash it several times with DPBS containing antibiotics, cut it into small pieces of about 5mm, add 10ml Dispase II separation enzyme, digest at 4°C for 17h, and obtain the cultured Umbilical cord mesenchymal cells.

[0037] 2. Plating of trophoblast cells: Prepare the temperature-sensitive hydrogel on the culture plate for 24 hours, and use the cultured umbilical cord mesenchymal cells as trophoblast cells by 2*10 4 / cm 2 , pre-seeded onto thermosensitive hydrogel-coated culture plates.

[0038] 3. Epidermal tissue pretreatment 2: Remove the epidermal tissue with Dispase II separation enzyme and wash it with DPBS 3 times, use toothed curved tweezers to peel off the epidermal layer, transfer the epidermal layer to a centrifuge tube, wash with DPBS, cut it into piece...

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Abstract

A method for culturing autologous tissue engineered epidermis, using umbilical cord mesenchymal cells as trophoblast cells. The advantage of the present invention is that by using umbilical cord mesenchymal cells as trophoblast cells, and at the same time cooperating with the serum-free medium of the present invention, there is no need to replace it with a differentiation medium, reducing the consumption of manpower, financial resources and time; Prepared with glue to avoid secondary digestion of the epidermal tissue by enzymes, reduce damage to cell activity, and accelerate the peeling speed between the epidermal tissue and the culture plate.

Description

technical field [0001] The invention relates to the technical field of cell culture, in particular to a method for culturing autologous tissue engineered epidermis and a culture medium thereof. Background technique [0002] At present, tissue-engineered epidermis is mainly used for a series of skin diseases such as burns, scalds, and vitiligo, and has a wide range of applications. Therefore, there is great concern on how to improve the quality and culture speed of tissue-engineered epidermis. There are two technical difficulties in the existing culture technology: [0003] First, about the culture of epidermal cells. In 1975, Rheinwald and Green invented the method of culturing mouse embryonic fibroblast cells (3T3) as trophoblast cells. In order to solve the safety problems caused by animal-derived cell lines, many people also used autologous fibroblasts as trophoblast cells. Cultivate; This method mainly has following four defects: a) the safety problem of animal source ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/071
CPCC12N5/0625C12N5/0626C12N2502/137
Inventor 柴勋
Owner SHANGHAI BIOMED UNION BIOTECHNOLOGY CO LTD
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