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Stem cell serum-free culture medium and preparation method and application thereof

A serum-free medium and stem cell technology, applied in the field of stem cell serum-free medium and its preparation, can solve the problem that mesenchymal stem cell-related products cannot meet clinical treatment requirements, high risk of allogeneic immune rejection, and unstable cultured cell quality, etc. It can protect and maintain growth and proliferation in vitro, improve the speed of in vitro culture, and avoid unfavorable factors.

Inactive Publication Date: 2021-08-10
KYBIOSTEM CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the composition of FBS is still unclear. The quality is closely related to the age, sex, and health of the donor. The content of different batches of substances varies greatly, which will lead to unstable quality of cultured cells and residual heterologous proteins. Clinical application brings a very high risk of alloimmune rejection
Based on the above reasons, the use of FBS will cause mesenchymal stem cell-related products to fail to meet clinical treatment requirements

Method used

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  • Stem cell serum-free culture medium and preparation method and application thereof
  • Stem cell serum-free culture medium and preparation method and application thereof
  • Stem cell serum-free culture medium and preparation method and application thereof

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preparation example Construction

[0050] In a second aspect, the present invention provides a method for preparing a serum-free medium for stem cells, comprising the following steps: adding platelet lysate, transferrin solution and human insulin solution to the basal medium to prepare the serum-free medium for stem cells.

[0051] The preparation method of the stem cell serum-free medium provided by the invention is simple and convenient.

[0052] As a further technical solution, before adding the transferrin solution and the human insulin solution to the basal medium, it also includes the step of filtering the transferrin solution and the human insulin solution to remove the impurities in the transferrin solution and the human insulin solution. of insoluble substances to obtain a homogeneously dissolved solution.

[0053] The filtering includes filtering with a filter membrane of 0.20 μm to 0.25 μm;

[0054] Preferably, a 0.22 μm filter membrane is used for filtration.

[0055] In the third aspect, the pres...

Embodiment 1

[0064] A stem cell serum-free medium, consisting of α-MEM medium and additional components, the additional components are platelet lysate, recombinant transferrin and recombinant human insulin, wherein the volume of platelet lysate in the stem cell serum-free medium The percentage is 5%, the concentration of recombinant transferrin is 5 μg / mL, and the concentration of recombinant human insulin is 5 μg / mL.

[0065] The preparation method is as follows:

[0066] Dissolve recombinant human insulin with 10mM hydrochloric acid to a concentration of 1mg / mL, pass through a 0.22 micron filter membrane, and add it to the basal medium at a final concentration of 5μg / mL; dissolve recombinant transferrin with sterile water to a concentration of 1mg / mL, after passing through a 0.22 micron filter membrane, it was added to the basal medium at a final concentration of 5 μg / mL; the platelet lysate was added to the above medium, and the volume ratio of the platelet lysate to the above medium w...

Embodiment 2

[0068] A stem cell serum-free medium, consisting of α-MEM medium and additional components, the additional components are platelet lysate, recombinant transferrin and recombinant human insulin, wherein the volume of platelet lysate in the stem cell serum-free medium The percentage is 5%, the concentration of recombinant transferrin is 10 μg / mL, and the concentration of recombinant human insulin is 10 μg / mL.

[0069] The preparation method is as follows:

[0070] Dissolve recombinant human insulin with 10mM hydrochloric acid to a concentration of 1mg / mL, pass through a 0.22 micron filter membrane, and add it to the basal medium at a final concentration of 10μg / mL; dissolve recombinant transferrin with sterile water to a concentration of 1mg / mL, after passing through a 0.22 micron filter membrane, it was added to the basal medium at a final concentration of 10 μg / mL; the platelet lysate was added to the above medium, and the volume ratio of the platelet lysate to the above medi...

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Abstract

The invention provides a stem cell serum-free culture medium and a preparation method and application thereof, and relates to the technical field of biology. The stem cell serum-free culture medium is composed of the basic culture medium and the additive component, the additive component is composed of platelet lysate, transferrin and human insulin, the stem cell serum-free culture medium is definite in component and low in cost, all the components cooperate with one another, proliferation of stem cells can be promoted, the in-vitro culture speed can be increased, serum-free addition is achieved, the safety is high, adverse factors caused by a serum-containing culture medium are avoided, and the culture medium is suitable for clinical application.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a serum-free culture medium for stem cells and its preparation method and application. Background technique [0002] Stem cells refer to the group of cells that maintain an undifferentiated and immature state during the development of the organism. During in vitro culture, stem cells can continuously regenerate, and under appropriate conditions, they can differentiate into various tissue cell types. The regenerative capacity of certain tissues in the body is directly related to the action of stem cells. In general, stem cells can be divided into three types according to their development, embryonic stem cells, induced pluripotent stem cells, and adult stem cells. [0003] As commonly used cells in regenerative medicine, stem cells are low in normal adult tissues and need to be expanded and cultured in vitro. Therefore, in vitro expansion of stem cells is a necessary condition to pr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071
CPCC12N5/0602C12N2500/90C12N2500/84C12N2500/25
Inventor 刘彦彦雷童李晓梅杜宏武
Owner KYBIOSTEM CO LTD
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