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Culture medium for in-vitro amplification of autologous human hair follicle mesenchymal stem cells and culture method thereof

A technology of mesenchymal stem cells and in vitro expansion, applied in the direction of cell culture active agent, culture process, tissue culture, etc., can solve the hidden dangers of autologous hair follicle mesenchymal stem cell, mycoplasma, soft virus infection, cross transmission and other problems, to achieve The effect of maintaining clonal proliferation ability and multi-directional differentiation potential, fast proliferation rate, and ensuring safety

Pending Publication Date: 2020-10-30
李鹏东 +1
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  • Summary
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  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The traditional culture of autologous human hair follicle mesenchymal stem cells is mainly fetal bovine serum or allogeneic human platelet lysates, which will bring a large amount of heterologous or allogeneic substances during the culture process, which makes autologous hair follicle mesenchymal stem cells clinically applicable. pose a great security risk
Clinical application The addition of animal components in autologous hair follicle mesenchymal culture will cause various animal viruses, mycoplasma, and soft viruses to infect the human body, which means that there are unknown potential risks when cells cultured in vitro are applied to the human body
Moreover, the addition of allogeneic human serum will bring cross-transmission of infectious diseases such as hepatitis B, tuberculosis, and syphilis
At the same time, when a small amount of heterologous protein enters the human body, it will cause a strong allergic reaction, even life-threatening

Method used

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  • Culture medium for in-vitro amplification of autologous human hair follicle mesenchymal stem cells and culture method thereof
  • Culture medium for in-vitro amplification of autologous human hair follicle mesenchymal stem cells and culture method thereof
  • Culture medium for in-vitro amplification of autologous human hair follicle mesenchymal stem cells and culture method thereof

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preparation example Construction

[0028] The present invention has no special limitation on the preparation method of the culture medium, and the preparation method of the culture medium known in the art can be adopted.

[0029] The present invention provides a method for in vitro expansion of autologous human hair follicle mesenchymal stem cells, comprising the following steps:

[0030] 1) Pre-cultivating autologous hair follicles in hair follicle medium to obtain autologous hair follicle mesenchymal stem cells;

[0031] The hair follicle culture medium is based on DMEM / F-12 medium, which contains 5% autologous serum and 1% penicillin-streptomycin in percentage by volume;

[0032] 2) Expanding and culturing the autologous hair follicle mesenchymal stem cells described in step 1) in the culture medium to obtain an autologous hair follicle mesenchymal stem cell bank.

[0033] In the invention, autologous hair follicles are pre-cultured in hair follicle medium to obtain autologous hair follicle mesenchymal stem...

Embodiment 1

[0038] A medium for expanding autologous human hair follicle mesenchymal stem cells in vitro, prepare sterilized DMEM / F-12 medium in advance, and then weigh autologous serum, bFGF, penicillin and streptomycin. For the preparation of autologous serum, the venous blood of the volunteers was left standing at 4°C for 1 hour, and the whole blood was centrifuged at 3000 rpm for 15 minutes to collect the supernatant, which was filtered through a 0.22 μm filter membrane , obtained autologous serum. Autologous serum, bFGF and penicillin-streptomycin were added to DMEM / F-12 medium according to the volume percentage of 5% autologous serum, 10ng / ml bFGF and 1% penicillin-streptomycin in turn, and filtered through microfiltration , to obtain the expansion medium.

Embodiment 2

[0040] A medium for expanding autologous human hair follicle mesenchymal stem cells in vitro, prepare sterilized DMEM / F-12 medium in advance, and then weigh autologous serum, bFGF, penicillin and streptomycin. For the preparation of autologous serum, the venous blood of volunteers was left standing at 4°C for 1.5h, and the whole blood was centrifuged at 3200rpm for 14min, the supernatant was collected, and the supernatant was filtered through a 0.22μm filter membrane , obtained autologous serum. Add autologous serum, bFGF, and penicillin-streptomycin to DMEM / F-12 medium in sequence according to the volume percentage of 5.2% autologous serum, 11ng / ml bFGF, and 0.8% penicillin-streptomycin, and filter through microfiltration Sterilize to obtain the expansion medium.

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Abstract

The invention provides a culture medium for in-vitro amplification of autologous human hair follicle mesenchymal stem cells and a culture method of the culture medium, and belongs to the technical field of cell culture. The culture medium for in-vitro amplification of the autologous human hair follicle mesenchymal stem cells takes DMEM / F-12 as a basic culture medium and is prepared from the following components in percentage by volume: 4 to 6 percent of autologous serum, 9 to 11ng / ml of bFGF and 0.8 to 1.2 percent of penicillin-streptomycin. According to the culture method, human autoserum isused as an additive to prepare an amplification culture medium, and in-vitro large-scale amplification of autologous human hair follicle mesenchymal stem cells is carried out. Compared with the traditional mesenchymal stem cells, the hair follicle mesenchymal stem cells cultured by the method have the advantages that other animal serum or allogeneic blood products are not required to be added, thesafety is high, the allogeneic disease propagation risk is avoided, the cell proliferation rate is high, and the characteristics of the mesenchymal stem cells can still be maintained after multiple passages.

Description

technical field [0001] The invention belongs to the technical field of cell culture, and in particular relates to a medium for expanding autologous human hair follicle mesenchymal stem cells in vitro and a culture method thereof. Background technique [0002] The discovery of stem cells and the establishment of transplantation technology have brought new vitality and hope to the treatment of diseases that are considered "incurable" in traditional medicine. Stem cells also play a huge role in medical beauty and anti-aging. Mesenchymal stem cells from multiple sources are currently the most widely used and safest stem cells. Mesenchymal stem cells derived from hair follicles are considered to be ideal stem cells at present, and have the following advantages: 1) High safety and no tumorigenicity, everyone can save their own hair follicle mesenchymal stem cells, and use them when needed after preservation In the body, there is no risk of disease transmission, and it is a reser...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775
CPCC12N5/0666C12N2500/84C12N2501/115
Inventor 李鹏东钟祥凤
Owner 李鹏东
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