Bionic three-dimensional DPCs independent co-culture system and construction method thereof

A technology of co-culture system and construction method, applied in the field of three-dimensional cell culture, can solve problems such as the research of DPCs microenvironment

Active Publication Date: 2019-12-17
NANFANG HOSPITAL OF SOUTHERN MEDICAL UNIV
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Problems solved by technology

After a large amount of literature research, we found that most of the current research is limited to the improvement of DPCs culture methods, and has not conducted in-depth research on the DPCs microenvironment, which is an important factor affecting the maintenance of normal biological functions of DPCs in vitro

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  • Bionic three-dimensional DPCs independent co-culture system and construction method thereof
  • Bionic three-dimensional DPCs independent co-culture system and construction method thereof
  • Bionic three-dimensional DPCs independent co-culture system and construction method thereof

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Embodiment Construction

[0050] The technical solutions of the present invention will be further described below through specific embodiments.

[0051] Unless otherwise specified, various raw materials and equipment of the present invention can be purchased commercially, or prepared according to conventional methods in the technical field.

[0052] The whole process of preparation is strictly required to be aseptic, and all the instruments used need to be sterilized by high temperature, high pressure or gas.

[0053] The construction method of the bionic three-dimensional DPCs independent culture system of this embodiment is as follows:

[0054] (1) Cell source: Using fresh hair follicle units from patients undergoing FUE hair transplantation, wash the hair follicle units extracted during the operation with PBS, cut off the hair follicles at 1-2 mm from the lower edge of the sebaceous gland under a stereoscope, take the lower part of the hair follicles, and pass through After digestion with 0.1% neut...

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Abstract

The invention discloses a bionic three-dimensional DPCs independent co-culture system and a construction method thereof. The construction method comprises the following steps: preparing single-cell suspensions of DPCs, DSCs and HGMCs; providing Matrigel, adding the DPCs single-cell suspension into the Matrigel, and carrying out resuspension to form a Matrigel-DP cell suspension; enabling the Matrigel-DP cell suspension to be adhered to a microporous membrane of a Transwell cell, so as to prepare a three-dimensional DPCs culture system; inoculating HGMCs in an upper chamber of the Transwell cell of the three-dimensional DPCs culture system, simultaneously inoculating DSCs in a lower chamber, and then adding a culture medium to prepare the bionic three-dimensional DPCs independent co-culturesystem. The in-vivo microenvironment of the DPCs can be effectively simulated, the difference between in-vitro cultured DPCs and in-vivo normal DPCs is reduced, and in-vitro long-time maintenance ofbiological functions of the DPCs is realized.

Description

technical field [0001] The invention relates to the technical field of three-dimensional cell culture, in particular to a method for constructing a bionic three-dimensional DPCs independent co-culture system and a method for cultivating the bionic three-dimensional DPCs independent co-culture system. Background technique [0002] Hair follicles contain dermal components (dermal papilla cells (DPCs), dermal sheath cells (DSCs)) and epidermal components (hair matrix cells (HGMCs), hair follicle stem cells, inner root sheath cells and outer root sheath cells). Among them, DPCs in the dermis play a key role in hair follicle morphogenesis and cycle, and are the core seed cells in hair follicle tissue engineering. The research on the biological function of DPCs is the key research direction of hair follicle tissue engineering. Although DPCs are similar to fibroblasts in origin, there are essential differences between them. First, DPCs express their own specific markers (ALP, Ve...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071
CPCC12N5/0627C12N2502/092C12N2513/00C12N2533/90
Inventor 苗勇胡志奇杜丽娟
Owner NANFANG HOSPITAL OF SOUTHERN MEDICAL UNIV
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