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A kind of preparation method of hair follicle stem cell active factor freeze-dried powder for treating hair loss

A technology for hair follicle stem cells and active factors, which is applied in the field of preparation of lyophilized powder of hair follicle stem cell active factors for the treatment of hair loss, can solve the problems of short storage time, unfavorable transportation, and reduced activity of stem cell culture medium, and achieves protection of activity and stability. Convenient, Bioactive Variety of Effects

Active Publication Date: 2022-08-02
安徽科门生物科技有限公司
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Problems solved by technology

[0007] The hair follicle stem cell active factor freeze-dried powder is used to replace the hair follicle stem cell factor solution to solve the following problem: some of the cytokines secreted by stem cells will decrease or lose their activity at normal temperature or under the condition of temperature change, and the expected effect cannot be achieved; Stem cell culture medium in liquid state has a short storage time in the greenhouse, which is not conducive to transportation

Method used

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  • A kind of preparation method of hair follicle stem cell active factor freeze-dried powder for treating hair loss
  • A kind of preparation method of hair follicle stem cell active factor freeze-dried powder for treating hair loss
  • A kind of preparation method of hair follicle stem cell active factor freeze-dried powder for treating hair loss

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preparation example Construction

[0037] see figure 1 As shown, the preparation method of hair follicle stem cell active factor freeze-dried powder provided by the present invention comprises the following steps:

[0038] S1. Acquisition of hair follicle stem cells: Under aseptic conditions, the roots of hair are cut to obtain hair follicle tissue, and then the hair follicle tissue is washed in DPBS containing double antibodies, then cut into pieces, and inoculated into the primary medium for the original Subculture, then digest and purify to obtain hair follicle stem cells;

[0039] S2, the acquisition of hair follicle stem cell active factors: the hair follicle stem cells are connected for subculture, and then enriched, centrifuged, and concentrated by ultrafiltration to obtain the hair follicle stem cell active factor concentrate;

[0040] S3. Mixing and freeze-drying: after mixing the hair follicle stem cell active factor concentrate and the auxiliary solution in a mixer, a mixed solution is obtained; aft...

Embodiment 1-1

[0047] Acquisition of hair follicle stem cells: Under aseptic conditions, the root of the hair is cut to obtain the hair follicle tissue, and then the hair follicle tissue is washed in DPBS containing penicillin and streptomycin, and then chopped (0.8mm). 3 ), and inoculated in the primary culture medium for primary culture, then digested and purified to obtain hair follicle stem cells;

[0048] Among them, the primary culture medium is HG-DMEM medium containing 10% fetal bovine serum by volume, and the medium is changed every 3 days in the primary culture; the specific operation of digestion is to obtain hair follicles when the cell density is 70-75%. tissue cells, then add neutral protease, hair follicle tissue cells and PBS buffer into a centrifuge tube to seal, and place it on a shaker at 37°C at 100r / min to digest for 1.5h, centrifuge at 2000rpm and discard the supernatant, and then use sterile PBS buffer The solution was rinsed twice to remove neutral protease, then 0.01...

Embodiment 1-2

[0050] Acquisition of hair follicle stem cells: Under aseptic conditions, the root of the hair is cut to obtain the hair follicle tissue, and then the hair follicle tissue is washed in DPBS containing penicillin and streptomycin, and then chopped (0.8mm). 3 ), and inoculated in the primary culture medium for primary culture, then digested and purified to obtain hair follicle stem cells;

[0051] Among them, the primary culture medium is HG-DMEM medium containing 10% fetal bovine serum by volume, and the medium is changed every 3 days in the primary culture; the specific operation of digestion is to obtain hair follicles when the cell density is 75-85%. Tissue cells, then add neutral protease, hair follicle tissue cells and PBS buffer into a centrifuge tube to seal, and place it on a shaker at 37°C at 100 r / min to digest for 2 hours. After centrifugation at 2000 rpm, discard the supernatant, and then use sterile PBS buffer. Rinse twice to remove neutral protease, then add 0.125...

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Abstract

The invention discloses a preparation method of freeze-dried powder of active factor of hair follicle stem cells for treating hair loss, belonging to the technical field of cytokines. And the preparation method includes the following steps: S1, acquisition of hair follicle stem cells; S2, acquisition of hair follicle stem cell active factors: inserting hair follicle stem cells into an expansion medium for subculture, and then enriching, centrifuging, ultrafiltration and concentrating, The hair follicle stem cell active factor concentrate is obtained; S3, mixing and freeze-drying: after mixing the hair follicle stem cell active factor concentrate and the auxiliary solution in a mixing box, a mixed solution is obtained; after the mixed solution is lyophilized for a second time, a treatment for hair loss is obtained. Freeze-dried powder of hair follicle stem cell active factor. Moreover, the invention replaces the hair follicle stem cell factor solution with the hair follicle stem cell active factor freeze-dried powder, so as to solve the problem that the hair follicle stem cell culture solution in the liquid state has a short storage time in the greenhouse and is not conducive to transportation.

Description

technical field [0001] The invention belongs to the technical field of cytokines, and in particular relates to a preparation method of freeze-dried powder of active factors for hair follicle stem cells for treating hair loss. Background technique [0002] In recent years, affected by various factors such as environment, work pressure, genetics, etc., the number of people with hair loss has been increasing. Hair loss is caused by many internal factors, such as aging or disturbances in hormonal balance, as well as various diseases and severe burns or wounds. For the treatment of hair follicle-related diseases such as alopecia and alopecia areata, the current conventional treatment methods include gene therapy, drug therapy and hair transplantation. There are drawbacks to the above treatment methods. For example, medication for hair loss and alopecia areata can lead to recurrence when the medication is discontinued, and if continued use can lead to accumulation of side effect...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K38/19A61K9/19A61K35/36A61P17/14C12N5/074C12P21/02
CPCA61K35/36A61K9/19A61P17/14C12N5/0628C12P21/02C12N2509/10C12N2509/00C12N2500/25C12N2501/39C12N2501/11
Inventor 张正亮
Owner 安徽科门生物科技有限公司
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