Preparation method of cell suspension for promoting hair regeneration and preventing alopecia

A technology of cell suspension and hair, which is applied in the field of cell culture, can solve problems such as insufficient donor sites, affecting surgical results, and limited sources, and achieve the effects of high efficiency, good cell activity, and strong regenerative ability

Inactive Publication Date: 2020-01-17
JINAN PANSHENG BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Hair transplantation uses the occipital region as the hair donor area, and the source of hair is limited. Therefore, for patients with large areas of baldness, there are often problems such as insufficient donor area, scarring of the donor area, and unstable transplant survival rate, which obviously affect the surgical effect.

Method used

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  • Preparation method of cell suspension for promoting hair regeneration and preventing alopecia
  • Preparation method of cell suspension for promoting hair regeneration and preventing alopecia
  • Preparation method of cell suspension for promoting hair regeneration and preventing alopecia

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Embodiment 1 prepares the cell suspension that promotes hair regeneration and prevents hair loss

[0033] 1. Materials and methods

[0034] (1) Treatment of scalp tissue: cut off the hair outside the scalp and the fat and connective tissue in the subcutaneous tissue, disinfect the collected fresh scalp tissue samples with alcohol for 3 minutes, and then treat them twice with PBS containing 2 times double antibody, each 3 minutes to achieve the purpose of sterilization. Drain the excess liquid from the processed tissue, weigh and make a record, and spread the tissue on a 100mm cell culture dish. Cut the tissue into uniform minces with a scalpel.

[0035] (2) Enzyme digestion: add the chopped tissue into the enzyme digestion solution (20ml enzyme digestion solution for 1g tissue, containing 2.5g / L type Ⅰ collagenase and 2.5g / L dissociating enzyme), and mix well. Digest in a 37°C water bath with 80×g shaking for 60 minutes. Then add 0.25% trypsin solution (5ml for 1g t...

Embodiment 2

[0041]Such as figure 1 As shown, Figure A is the prepared scalp epidermal cells, and Figure B is the cultured dermal papilla cells. Example 2 Experiment on hair regeneration effect of immunized mice injected with cell suspension on the back

[0042] 1. Materials and methods

[0043] (1) Scalp epidermal cells and dermal papilla cell suspension injection

[0044] After cultivating qualified human scalp epidermal cells and dermal papilla cells, according to a certain ratio (in this implementation case, the number of scalp epidermal cells is 1 × 10 6 , the passage is P3; the number of dermal papilla cells is 2×10 6 , the passage is P4) mixed with 75 microliters of F12 medium to prepare a cell suspension. Inject four injections into the back of immunodeficient mice.

[0045] (2) Analysis of hair regeneration

[0046] Twelve weeks after the injection, check the hair follicle regeneration on the mice with a dissecting optical microscope, excise and collect the regenerated skin ...

Embodiment 3

[0050] Example 3 Clinical Trial of Autologous Scalp Tissue-derived Cell Suspension Injection

[0051] 1. Epidermal cell and dermal cell suspension injection

[0052] Select 20 androgenetic alopecia patients aged 30-50 with a course of disease within 2 years, and other healthy men with physical indicators, and sign the informed consent form with the consent of the patients before use. Use a scalpel to take 2*2 cm2 scalp tissue from the patient's occiput, and culture human scalp epidermal cells and dermal papilla cells according to the above-mentioned experimental method. Qualified human scalp hair follicle cells and dermal papilla cells after cultivation, according to a certain ratio (the number of scalp epidermal cells in this implementation case is 2 × 10 6 , the passage is P3; the number of dermal papilla cells is 5×10 6 , the passage is P4) mixed with 75 microliters of F12 medium to prepare a cell suspension. Routinely disinfect the skin of the patient, inject it at mult...

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PUM

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Abstract

The invention discloses a preparation method of a cell suspension for promoting hair regeneration and preventing alopecia. According to the method, scalp epidermal cells and dermal papilla cells are separated from scalp tissues, and the scalp epidermal cells and the dermal papilla cells are subjected to multiplication culture to prepare the cell suspension for preparing an injection for treating androgenic alopecia, and the injection is derived from patients without immune rejection reaction, has good cell activity, strong regeneration capability and high effective rate for treating the androgenic alopecia; and moreover, the injection can be prepared only by taking a small amount of scalp tissues, and the transplantation efficiency is high.

Description

technical field [0001] The invention relates to a preparation method of a cell suspension for promoting hair regeneration and preventing hair loss, and belongs to the field of cell culture. Background technique [0002] Alopecia is a relatively common disease in clinical dermatology. At present, the incidence rate is gradually increasing and showing a younger trend, which plagues the lives of modern people, and thus has received more and more attention. The hair that falls out normally is the hair in the catagen and telogen phase. Since the hair entering the catagen and the newly entering the growth phase are constantly in a dynamic balance, it can maintain a normal amount of hair. If the hair falls out abnormally or excessively, it belongs to abnormal hair loss, and there are many reasons. There are hereditary, pathological, and bad living habits that cause hair loss. [0003] Androgenetic alopecia (also known as male pattern alopecia, MA) is the most common progressive a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071A61K35/36A61P17/14
CPCC12N5/0625A61K35/36A61K9/0019A61P17/14C12N2509/00C12N2501/727
Inventor 张平邢志青吴训伟张甜甜王杰
Owner JINAN PANSHENG BIOTECH
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