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Artificial hair follicle, preparation method therefor and use thereof

A hair follicle and artificial technology, applied in the biological field, can solve the problems of affecting the survival rate of hair follicles, mechanical damage of hair follicles, and the inability of patients with hair loss to have a good treatment effect, and achieve the effect of improving the efficiency of hair follicle formation and the induction efficiency.

Active Publication Date: 2019-08-06
SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the method of hair follicle transplantation is to take the hair follicles from the back of the occiput, and then transplant the hair follicles to the same person's forehead where the hair is sparse. It is effective for patients with a small amount of hair loss, but it cannot have a good therapeutic effect for patients with large areas of hair loss
At the same time, hair follicle extraction will cause mechanical damage to hair follicles, which will affect the survival rate of hair follicles.

Method used

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  • Artificial hair follicle, preparation method therefor and use thereof
  • Artificial hair follicle, preparation method therefor and use thereof
  • Artificial hair follicle, preparation method therefor and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] 1. Cell provider's peripheral blood cell preparation, induction and detection:

[0042] 30ml of peripheral blood was isolated from the cell donor by venous blood draw, and mononuclear (MNC) cells were obtained by Ficoll-Hypaque density gradient separation. MNC cells were cultured in α-MEM medium containing 10% FBS and 10 ng / ml IL-7 or 10 ng / ml G-CSF, GM-CSF, IL-3 and IL-6 for 5 days. These cells are then used to generate induced pluripotent stem cells, the specific steps are as follows, 1-2×10 4 Each MNC was placed in 1 well of a 96-well plate, and was infected with a CytoTune-iPS reprogramming kit (DNAVEC, Tsukuba, Japan) encoding OCT4, SOX2, KLF4, cMYC F-deficient Sendai virus vector (Sev / ΔF) , where the multiplicity of infection (MOI) for each factor is 5-10. One day after infection, remove the culture medium by low-speed centrifugation, harvest the cells, and culture them in a six-well plate cultured with mouse embryonic fibroblasts (MEF) as the bottom, and cultur...

Embodiment 2

[0048] 1. Preparation of feeder-free culture for differentiation of iPS cells into skin organoids

[0049] This operation needs to be performed in a sterile biological safety cabinet. Similar to Matrigel, Geltrex matrix solidifies rapidly at room temperature (RT), thus requiring pre-frozen tips, racks, and tubes when new gels are aliquoted. Make 50, 100, and 200 µL aliquots and store them at -80 °C. Use Geltrex at a 1:100 dilution. Feeder-free iPSC culture only requires Geltrex as a surface coating agent, however for iPSC differentiation, the combination of Geltrex and ColI is more effective in inducing the differentiation of iPS cells into keratinocytes. For 60mm tissue culture dish Geltrex surface coating method is as follows. If larger dishes are to be used, adjust the volume of Geltrex solution accordingly. Remove 50 µL of Geltrex from the -80 °C freezer and place it on ice inside the biosafety cabinet. Add 5 mL of cooled sterile DMEM / F12 to a 15 mL centrifuge tube. ...

Embodiment 3

[0055] 1. Cell transplantation and hair formation and hair follicle isolation

[0056] Using induced pluripotent stem cells to form a hair follicle structure, the specific process is as follows: (1) culture the partially differentiated induced pluripotent stem cells obtained in Example 2, digest them with Accutase enzyme, and prepare a cell suspension with a total cell number of 2 million In PBS, centrifuge at 300g for 5min. (2) Take the centrifuged cells, discard the supernatant, add 50 microliters of Geltrex to the cell pellet, mix well, and wait for use. At the same time, add Wnt10b factor so that the concentration in Geltrex is 1 ug / ml. (3) The embryonic pig about 60 days pregnant is taken out from the abdominal cavity of the sow by laparotomy, but the pig fetus is not allowed to leave the mother's body, and a wound with an area of ​​about 7mm2 is made on the skin of its back with a puncher. (4) The Geltrex mixed with skin organoid cells prepared in step (2) was transplan...

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Abstract

The invention provides artificial hair follicle, a preparation method therefor and use thereof. The method comprises: transplanting human pluripotent stem cells into the skin of a heterologous embryoat the hair forming stage; continuously cultivating the embryo transplanted with the human pluripotent stem cells in a heterologous maternal body until the embryo is delivered; and isolating hair follicles in the skin of the embryo transplanted with the human pluripotent stem cells to obtain an artificial hair follicle. The artificial hair follicle obtained by the method according to the embodiment of the invention has a large number, the structural morphology is similar to that of the hair follicle in normal skin, and the artificial hair follicle can be used for hair follicle transplantationof different parts of the human body, such as scalp, eyebrow, beard, body hair, and the like; at the same time, the obtained artificial hair follicles are transplanted back to a pluripotent stem cellprovider without immunological rejection, and the safety is greatly improved.

Description

technical field [0001] The present invention relates to the field of biotechnology, in particular, the present invention relates to artificial hair follicles and their preparation methods and applications. Background technique [0002] The thinning of hair caused by hair loss has become an increasingly serious sub-health problem that plagues modern men (including some women). Although hair loss is not a serious physical health problem, hair loss will seriously affect the individual's psychological state, so it will be serious reduce the individual's quality of life. According to incomplete statistics, there are currently more than 200 million people in my country who suffer from hair loss to varying degrees, and this group of people in the world has exceeded 1 billion. Pathological abnormal hair loss, such as male baldness, is that under the action of certain factors, the hair follicles are in the resting period for a long time and cannot enter the growth cycle. Now there a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071A61F2/10
CPCC12N5/0698A61F2/10C12N2506/45A61B2017/00752
Inventor 王旭升吴耀炯牟丽莎曹楠
Owner SUN YAT SEN UNIV
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