Serum-free medium for stem cells

A serum-free medium and stem cell technology, applied in the field of stem cell culture in vitro, can solve problems affecting cell survival rate and cell proliferation process, hematopoietic stem cell proliferation and differentiation, complex development and maturation process, etc., to reduce mortality and increase proliferation efficiency , improve the effect of activity

Pending Publication Date: 2019-04-09
广州和能生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] However, to make full use of umbilical cord blood stem cells, more methods are needed to increase the number of umbilical cord blood stem cells. At present, the most researched is the in vitro expansion of umbilical cord blood stem cells. However, human or animal serum is routinely added to the in vitro expansion, and serum contains Many uncertain substances (such as different antigens, antibodies, hormones and cytokines, etc.), in recent years, especially the virus infection in animals, the serum may carry some animal viruses, thi

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] The invention provides a technical solution: a serum-free medium for stem cells, characterized in that: taking the volume of the high-sugar DMEM basal medium with a glucose concentration of 3750 mg / L as a reference, the composition and content of the added components are respectively Ferritin, 7.5% by volume, insulin, 10mg / L, L-glutamine, 3mM, growth factor, 10ng / mL, BMP-5 antibody, 0.15ng / mL, lipid complex, 3mM and trace elements, 3mg / L, at the same time, the composition and content of the cytokine combination are: stem cell growth factor (SCF), 15ng / mL, granulocyte-macrophage colony-stimulating factor (GM-CSF), 10ng / mL, interleukin 3 (IL-3), 3ng / mL and interleukin 6 (IL-6) 8ng / mL, the DMEM basal medium and additives are mixed according to a specific volume ratio or content to prepare a culture solution, and within 14 days, the cells The combined factors were added to the culture medium in the order of stem cell growth factor (SCF), interleukin 3 (IL-3), interleukin 6 ...

Embodiment 2

[0020] The invention provides a technical solution: a serum-free medium for stem cells, characterized in that: taking the volume of the high-sugar DMEM basal medium with a glucose concentration of 4500 mg / L as a reference, the composition and content of the added components are respectively Ferritin, 10% by volume, insulin, 15mg / L, L-glutamine, 4mM, growth factor, 5ng / mL, BMP-5 antibody, 0.25ng / mL, lipid complex, 4mM, and trace elements, 5mg / L, at the same time, the composition and content of the cytokine combination are: stem cell growth factor (SCF), 20ng / mL, granulocyte-macrophage colony-stimulating factor (GM-CSF), 15ng / mL, interleukin 3 (IL-3), 5ng / mL and interleukin 6 (IL-6) 15ng / mL, the DMEM basal medium and additives are mixed according to a specific volume ratio or content to prepare a culture solution, and within 14 days, the cells The combined factors were added to the culture medium in the order of stem cell growth factor (SCF), interleukin 3 (IL-3), interleukin 6 ...

Embodiment 3

[0021] Embodiment three: a kind of stem cell serum-free culture medium, it is characterized in that: with the glucose concentration being the volume of the high-sugar DMEM basal medium of 3000mg / L as a benchmark, the composition and content of the added components are transferrin, respectively, 5% by volume, Insulin, 5mg / L, L-Glutamine, 2mM, Growth Factor, 5ng / mL, BMP-5 Antibody, 0.05ng / mL, Lipid Complex, 2mM and Trace Elements, 1mg / L , meanwhile, the composition and content of the cytokine combination are respectively: stem cell growth factor (SCF), 10ng / mL, granulocyte-macrophage colony-stimulating factor (GM-CSF), 5ng / mL, interleukin-3 (IL -3), 1ng / mL and interleukin 6 (IL-6) 1ng / mL, the DMEM basal medium and additives are mixed according to a specific volume ratio or content to prepare a culture solution, and within 14 days, the cells in the combined factor Add stem cell growth factor (SCF), interleukin 3 (IL-3), interleukin 6 (IL-6) and granulocyte-macrophage colony-stimu...

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Abstract

The invention discloses a serum-free medium for stem cells. The serum-free medium comprises a basic medium, an additive and a cytokine combination, wherein in terms of the volume of the basic medium as reference, the additive comprises transferrin in a volume ratio of 5%-10%, insulin with concentration of 5-15 mg/L, 2-4 mM of L-glutamine, growth factors with concentration of 5-15 ng/mL, BMP-5 antibody with concentration of 0.05-0.25 ng/mL, 2-4 mM of a lipid complex and trace elements with concentration of 1-5 mg/L. The serum-free animal-derived components are adopted, so that interference of many uncertain substances (such as different antigens, antibodies, hormone and cytokines) contained in serum on in-vitro expansion survival rate and the cell proliferation process of hematopoietic stemcells is eliminated, the proliferation efficiency is increased, the quantity of high-activity hematopoietic stem cells is increased, use types and adding content of the cytokines in the in-vitro expansion process of the hematopoietic stem cells are adjusted, then the activity of the hematopoietic stem cells in in-vitro expansion is improved, and death rate of the cells can be reduced.

Description

technical field [0001] The invention relates to the technical field of in vitro culture of stem cells, in particular to a serum-free culture medium for stem cells. Background technique [0002] Bone marrow hematopoietic stem cell and adult peripheral blood hematopoietic stem cell transplantation can treat hematological tumors (such as leukemia, multiple myeloma, etc.), aplastic anemia, immunodeficiency diseases and genetic diseases. In recent years, due to the advances in basic research on cord blood hematopoietic stem cells, it has been found that cord blood contains more abundant hematopoietic stem cells than bone marrow, and its ability to grow and expand is richer and easier to obtain than bone marrow. It is safe and easy to collect and less prone to virus transmission. and graft-versus-host disease. In the past, umbilical cord blood was a waste, but now, umbilical cord blood stem cells can be preserved through cord blood banks, which increases the possibility of its ra...

Claims

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Application Information

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IPC IPC(8): C12N5/0789
CPCC12N5/0647C12N2501/155C12N2500/90C12N2501/10C12N2500/25C12N2500/36C12N2500/32
Inventor 吴家波周海军
Owner 广州和能生物科技有限公司
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