Multifunctional stem cell culture medium

A technology of pluripotent stem cells and culture medium, applied in the field of pluripotent stem cell culture medium, can solve problems such as unfavorable cell culture and affecting cell growth

Pending Publication Date: 2021-03-09
无锡华泰创新药技术研究院有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] There are certain defects in the traditional pluripotent stem cell culture medium. During the culture of cell culture medium, the cells are easily infected by external bacteria. Bacteria will affect the growth of cells, which is not conducive to the cultivation of cells.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] First take 150 μL / cm2 Corning BD Matrigel to the bottom side of DMEM / F12 cell culture medium, then take 10ng / mL sodium selenite, 20ng / mL recombinant transferrin, 20μg / mL insulin, 400μg / mL allicin, 40 μg / mL streptomycin, the concentration of glucose is 10 mmol / L, the concentration of vitamin C is 6 μg / mL, the concentration of basic fibroblast growth factor is 10 ng / mL, and the concentration of transforming growth factor beta1 is 1 ng / mL .

[0029] Then sodium selenite, recombinant transferrin, insulin, allicin, streptomycin, glucose, vitamin C, basic fibroblast growth factor L and transforming growth factor beta1 were first mixed, and then added to DMEM / F12 cell culture medium Inside.

[0030] The osmotic pressure of the DMEM / F12 cell culture medium was adjusted to 300 mosmol / L, and the volume was constant to obtain a pluripotent stem cell culture medium.

Embodiment 2

[0032] First take 160 μL / cm2 Corning BD Matrigel to the bottom side of DMEM / F12 cell culture medium, then take 12ng / mL sodium selenite, 50ng / mL recombinant transferrin, 22μg / mL insulin, 440μg / mL allicin, 44 μg / mL streptomycin, the concentration of glucose is 12mmol / L, the concentration of vitamin C is 7μg / mL, the concentration of basic fibroblast growth factor is 14ng / mL, the concentration of transforming growth factor beta1 is 1.2ng / mL mL.

[0033] Then sodium selenite, recombinant transferrin, insulin, allicin, streptomycin, glucose, vitamin C, basic fibroblast growth factor L and transforming growth factor beta1 were first mixed, and then added to DMEM / F12 cell culture medium Inside.

[0034] The osmotic pressure of the DMEM / F12 cell culture medium was adjusted to 300 mosmol / L, and the volume was constant to obtain a pluripotent stem cell culture medium.

Embodiment 3

[0036] First take 170μL / cm2 of Corning BD Matrigel to the bottom side of DMEM / F12 cell culture medium, then take 14ng / mL sodium selenite, 80ng / mL recombinant transferrin, 24μg / mL insulin, 480μg / mL allicin, 48 μg / mL streptomycin, the concentration of glucose is 14 mmol / L, the concentration of vitamin C is 7.5 μg / mL, the concentration of basic fibroblast growth factor is 18 ng / mL, and the concentration of transforming growth factor beta1 is 1.4 ng / mL.

[0037] Then sodium selenite, recombinant transferrin, insulin, allicin, streptomycin, glucose, vitamin C, basic fibroblast growth factor L and transforming growth factor beta1 were first mixed, and then added to DMEM / F12 cell culture medium Inside.

[0038]The osmotic pressure of the DMEM / F12 cell culture medium was adjusted to 300 mosmol / L, and the volume was constant to obtain a pluripotent stem cell culture medium.

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Abstract

The invention discloses a multifunctional stem cell culture medium, and belongs to the technical field of bioengineering. The multifunctional stem cell culture medium comprises a basic culture mediumand the following added components: 150-200 microliters / cm<2> of Kangning BD Matrigel, 10-20 nanograms / mL of sodium selenite, 20-200 nanograms / mL of recombinant transferrin, 20-30 micrograms / mL of insulin, 400-600 micrograms / mL of allicin and 40-60 micrograms / mL of streptomycin. Due to adoption of the Kangning BD Matrigel, the Matrigel is polymerized to form a three-dimensional matrix with biological activity, the structure, the composition, the physical properties and the functions of an in-vivo cell basement membrane are simulated, and culture and differentiation of in-vitro cells are facilitated; under the action of the recombinant transferrin, the insulin and the like, normal proliferation of cells can be ensured, and the possibility of pathogen contamination is reduced; and due to combined use of the allicin and the streptomycin, the inhibition and killing functions of the allicin to bacteria can be remarkably improved, the cell culture medium disclosed by the invention has goodinhibition and killing functions to bacteria, growth and proliferation of cells are facilitated, and the possibility of bacterial infection in the cell culture medium is reduced.

Description

technical field [0001] The invention relates to the technical field of bioengineering, in particular to a culture medium for pluripotent stem cells. Background technique [0002] Pluripotent stem cells (PSCs) are a type of pluripotent cells with self-renewal and self-replication capabilities. Under certain conditions, it can differentiate into a variety of APSC pluripotent cells, and pluripotent stem cells (Ps) have the potential to differentiate into a variety of cell tissues. The human iPSC culture process has gone through three stages: using feeder cell culture, feeder-free cell culture, and animal-derived feeder-free cell culture. Feeder layer cells are usually derived from mouse embryonic fibroblasts (MEF), and the batch-to-batch differences, animal origin, various infectious origins, and component uncertainties affect the judgment of experimental research and further in-depth application research. [0003] The application of iPS cells to treat diseases is the ultimat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/074C12N5/0735
CPCC12N5/0606C12N5/0607C12N5/0696C12N2500/25C12N2500/30C12N2500/34C12N2500/38C12N2501/115C12N2501/148C12N2533/90
Inventor 陈晓栋尚倩雯贾善粉张胜超王旻贤马慧
Owner 无锡华泰创新药技术研究院有限公司
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