Primary sweat gland cell in-vitro separation method

A separation method and technology for sweat gland cells are applied in the field of in vitro culture and separation of primary sweat gland cells that can efficiently achieve rapid enrichment of primary sweat gland cells in vitro, can solve the problems of inability to achieve rapid enrichment of primary sweat gland cells and the like, and achieve beneficial activity. Maintain and improve the separation efficiency and the effect of high-efficiency separation

Pending Publication Date: 2022-03-01
GENERAL HOSPITAL OF PLA
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

[0004] The embodiment of the present invention provides a method for isolating primary sweat gland cells in vitro to solve the problem that the prior art cannot realize rapid enrichment of primary sweat gland cells in vitro

Method used

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  • Primary sweat gland cell in-vitro separation method
  • Primary sweat gland cell in-vitro separation method
  • Primary sweat gland cell in-vitro separation method

Examples

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Embodiment 1

[0072] (1) Take out the human breast tissue and put it into a 10cm cell culture dish, wash it three times with PBS added with 5% penicillin / streptomycin, then remove the fat part, cut the tissue into fine pieces, add 10mL 2mg / mL type II collagenase , stirred evenly and put into 37°C, 5% volume fraction of CO 2 Digest overnight in an incubator to obtain sweat gland tissue, specifically as figure 2 shown.

[0073] (2) Sweat gland conditioned medium configuration, the sweat gland conditioned medium is cell basal medium DMEM / F12 with a final concentration of 10% serum and 1% penicillin / streptomycin.

[0074] (3) Take out the 6cm cell culture dish, add 2mL IV collagen, put it in 37℃, 5% CO 2 Cover in the incubator for 15 minutes, take out the cell culture dish, suck off the IV collagen, wash it once with PBS, and then add the sweat gland conditioned medium configured in Example 1 to ensure that the sweat gland conditioned medium does not cover the bottom of the dish.

[0075] (...

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Abstract

The invention provides a primary sweat gland cell in-vitro separation method which comprises the following steps: coating a cell culture dish with collagen to obtain a coated cell culture dish; adding a sweat gland conditioned medium into the coated cell culture dish, and adding sweat gland tissues into the coated cell culture dish; and after the sweat gland tissue adheres to the wall and climbs out of the cells, realizing in-vitro high enrichment of primary sweat gland cells by using a differential digestion method. According to the method disclosed by the invention, the cell culture dish is coated with collagen to increase the adhesion of adherent cells, so that 90% of in-vitro sweat gland tissues can be quickly adhered to the wall within 8 hours and climb out of sweat gland cells within 24 hours. According to the improved method provided by the invention, the damage to the cells caused by secondary tissue digestion, high-strength mechanical blowing and beating and centrifugation involved in a traditional enzyme digestion method is avoided, the separation step is simplified, the separation efficiency of the primary sweat gland is improved, and the activity maintenance and in-vitro amplification of the primary sweat gland cells are facilitated.

Description

technical field [0001] The invention relates to the technical field of sweat gland cell culture, in particular to an in vitro culture and separation method for primary sweat gland cells that efficiently realizes rapid enrichment of primary sweat gland cells in vitro. Background technique [0002] Sweat glands are one of the most important appendages of the skin, which play a pivotal role in substance metabolism, water and electrolyte balance, and maintaining the homeostasis of the body. However, the regeneration ability of sweat glands is limited, and the cicatricial repair of the post-traumatic skin further destroys the structure and function of sweat glands, resulting in the inability of the skin to perspire normally and the imbalance of body temperature regulation, which brings great physical and psychological harm to patients. [0003] With the continuous deepening of research on skin wound repair, regenerative medicine centered on stem cell technology has been hailed as...

Claims

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Application Information

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IPC IPC(8): C12N5/071
CPCC12N5/0633C12N2501/998C12N2509/10C12N2509/00C12N2500/02C12N2501/70
Inventor 项江兵孙晓艳付小兵
Owner GENERAL HOSPITAL OF PLA
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