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Method for constructing androgenetic alopecia cell model

A technology of androgenetic and cell models, applied in the biological field, can solve the problem of not being able to truly simulate the shape of the dermal papilla, the lack of communication between cells and extracellular matrix, cell proliferation, migration, differentiation and in vivo differences in 2D androgenetic alopecia models Large and other problems, to achieve the effect of reducing modeling risk, shortening modeling time, and high reference value

Active Publication Date: 2021-11-09
NANFANG HOSPITAL OF SOUTHERN MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] The purpose of the present invention is to address the lack of communication between cells and extracellular matrix in the 2D androgenetic alopecia models existing in the prior art, and the two-dimensional growth of cell proliferation, migration, differentiation and in vivo differences are large, and it cannot be truly simulated. The problem of the shape of the dermal papilla bulb in the human body, providing a method for constructing a cell model of androgenetic alopecia

Method used

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  • Method for constructing androgenetic alopecia cell model
  • Method for constructing androgenetic alopecia cell model
  • Method for constructing androgenetic alopecia cell model

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Experimental program
Comparison scheme
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Embodiment 1

[0051] Step 1, obtaining dermal papilla bulbs, inoculating, culturing and subculture.

[0052] 1. Human hair follicles were taken from the bald (frontal) area of ​​a male patient undergoing hair transplantation;

[0053] 2. Put the hair follicles in DPBS and wash them three times, use microsurgical scissors to cut off the hair bulbs of the hair follicles under the body microscope, rinse the hair bulbs three times with DPBS, after exhausting the DPBS, add 3ml of 2mg / ml Collagenase digestion for 30 minutes, pipetting once every 10 minutes to obtain dermal papilla ball suspension;

[0054] 3. Centrifuge the dermal papilla ball suspension at 1000rpm for 5min, discard the supernatant, and resuspend with 1% (v / v) penicillin-streptomycin and DMEM medium containing 20% ​​(v / v) fetal bovine serum , inoculate the dermal papilla balls in a cell culture flask, add 1% (v / v) penicillin-streptomycin and DMEM medium containing 20% ​​(v / v) fetal calf serum, place at 37°C and 5% CO 2 culture...

Embodiment 2

[0069] The steps of this embodiment are the same as those of the above-mentioned embodiment 1, except that the concentration of dihydrotestosterone is 100 μM.

Embodiment 3

[0071] The steps of this embodiment are the same as those of the above-mentioned embodiment 1, except that the concentration of dihydrotestosterone is 1 μM.

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Abstract

The invention discloses a method for constructing an androgenetic alopecia cell model. The method comprises the following steps that hair follicle hair bulb parts of forehead parts of an androgenetic alopecia patient are taken, hair papilla of the hair follicle hair bulb parts is separated, and culturing is carried out; a carina region of an occipital part of the androgenetic alopecia patient is taken, outer root sheath cells of the carina region are separated, and culturing is carried out; 3D culture is carried out on the hair papilla cells obtained after culture treatment, so that the hair papilla cells are gathered to form hair papilla cell balls; and the hair papilla cell balls are inoculated into an upper chamber of a transwell culture dish, the outer root sheath cells are inoculated into a lower chamber of the pre-coated transwell culture dish, and dihydrotestosterone is added to obtain the androgenetic alopecia cell model. The androgenetic alopecia cell model is relatively stable, the number of the outer root sheath cells is reduced after successful modeling, the change trend of androgenetic alopecia related markers is consistent with that in vivo, and the androgenetic alopecia cell model has a relatively high reference value for testing androgenetic alopecia drugs or treatment means.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for constructing a cell model of androgenetic alopecia. Background technique [0002] Androgen alopecia (AGA), also known as male pattern baldness. Androgenetic alopecia is characterized by androgen-induced premature transition of hair follicles from anagen to catagen, as well as miniaturization of hair follicles. 1998) 235-248). AGA is a disease affected by multiple factors and has complex etiology. There are still many unknowns in the current etiology research. Previous studies have believed that one of the main causes of male hair loss is genetic factors. In recent years. With the development of molecular biology, the research on the pathogenesis of androgenetic alopecia has become more and more in-depth, and the theory that androgen may cause AGA has become more and more mature. Dermal papilla cells in androgenetic alopecia contain 5-alpha-reductase, which leads to ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071
CPCC12N5/0627C12N2513/00C12N2501/392C12N2509/00Y02A50/30
Inventor 苗勇胡志奇张钰凡杜丽娟
Owner NANFANG HOSPITAL OF SOUTHERN MEDICAL UNIV
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