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Mesenchymal stem cell serum-free medium and application thereof

A technology of serum-free medium and mesenchymal stem cells, which is applied in the field of serum-free medium for mesenchymal stem cells, can solve the problem of unrepeatable effects

Active Publication Date: 2021-04-06
YUNNAN KEY LAB OF PRIMATE BIOMEDICINE RES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The culture method involved in some domestic serum-free and heterologous protein-free mesenchymal stem cell serum-free medium patents cannot even repeat the effect it advertises

Method used

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  • Mesenchymal stem cell serum-free medium and application thereof
  • Mesenchymal stem cell serum-free medium and application thereof
  • Mesenchymal stem cell serum-free medium and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0071] This embodiment provides a serum-free medium for mesenchymal stem cells, each 1L of the medium consists of the following components:

[0072] IMDM 17.662g, L-alanyl-glutamine 2mM, chemical lipid 0.6v / v%, cholesterol 15mg, insulin 12.5mg, transferrin 25mg, recombinant human albumin 2g, hydrocortisone 500μg, dexamethasone 4μg, progesterone 5.66μg, putrescine 9mg, ascorbic acid 100mg, β-mercaptoethanol 75μM, soybean lecithin 10mg, zinc sulfate heptahydrate 2.5mg, lipoic acid 0.2mg, N-Acetyl-L-Cysteine ​​1mM, reduced glutathione Glycine 2mg, Taurine 5mg, Y-27632 5μM, Ethanolamine 2mg, b-FGF 20μg, EGF 20μg, PDGF-BB 10μg, IGF-1 15μg, TGF-β3 2μg, HGF 10μg, CTGF 2μg and VEGF 15μg; The amount is water for cell culture.

[0073] This embodiment further provides a method for configuring the above-mentioned serum-free medium for mesenchymal stem cells:

[0074] Dissolve IMDM powder in cell culture water, stir well, filter with 0.22um filter membrane, add recombinant human insulin...

Embodiment 2

[0076] This embodiment provides a serum-free medium for mesenchymal stem cells, each 1L of the medium consists of the following components:

[0077] IMDM 17.662g, L-alanyl-glutamine 2mM, chemical lipid 0.1v / v%, cholesterol 5mg, insulin 8mg, transferrin 10mg, recombinant human albumin 1g, hydrocortisone 0.1mg, dexamethasone 4μg, progesterone 1μg, putrescine 5mg, ascorbic acid 25mg, β-mercaptoethanol 50μM, soybean lecithin 2mg, zinc sulfate heptahydrate 1.25mg, lipoic acid 0.1mg, N-Acetyl-L-Cysteine ​​0.2mM, reduced glutathione Glycine 1mg, Taurine 2mg, Y-27632 2μM, Ethanolamine 1mg, b-FGF 5μg, EGF 5μg, PDGF-BB 2μg, IGF-1 10μg, TGF-β3 1μg, HGF 2μg, CTGF 1μg and VEGF 5μg; The amount is water for cell culture.

[0078] Its configuration method is the same as embodiment 1.

Embodiment 3

[0080] This embodiment provides a serum-free medium for mesenchymal stem cells, each 1L of the medium consists of the following components:

[0081] IMDM 17.662g, L-alanyl-glutamine 2mM, chemical lipid 1v / v%, cholesterol 30mg, insulin 25mg, transferrin 30mg, recombinant human albumin 5g, hydrocortisone 2mg, dexamethasone 20μg, Progesterone 10μg, putrescine 15mg, ascorbic acid 200mg, β-mercaptoethanol 100μM, soybean lecithin 20mg, zinc sulfate heptahydrate 2.5mg, lipoic acid 0.5mg, N-Acetyl-L-Cysteine ​​2mM, reduced glutathione 5mg , taurine 10mg, Y-27632 10μM, ethanolamine 5mg, b-FGF 40μg, EGF 50μg, PDGF-BB 20μg, IGF-1 40μg, TGF-β3 5μg, HGF 10μg, CTGF 5μg and VEGF 20μg; the balance was cells Cultivate water.

[0082] Its configuration method is the same as embodiment 1.

[0083] The same strain of P1 generation human umbilical cord mesenchymal stem cells was taken for testing, and three groups of repeated experiments were carried out in each of Examples 1-3, and the inoculat...

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Abstract

The invention relates to the technical field of cell culture, and particularly relates to a mesenchymal stem cell serum-free medium and application thereof. The mesenchymal stem cell serum-free medium comprises a basic medium and additive components, wherein the basic medium is an IMDM medium; the additive components comprise TGF-[beta]3 and ascorbic acid, the concentration of TGF-[beta]3 is 1-5 [mu]g / L, and the dosage ratio of ascorbic acid to TGF-[beta]3 is 25-200mg: 1-5[mu]g. The medium disclosed by the invention has a good cell adherence promoting effect, and the cultured cells are strong in multiplication capacity, complete in form and strong in secretion capacity. Compared with a traditional serum culture system and an existing serum-free system, the serum-free medium is clear in chemical components, free of human-derived or animal-derived protein and free of serum, and the instability of animal-derived components and batches is avoided.

Description

technical field [0001] The invention relates to the technical field of cell culture, in particular to a serum-free medium for mesenchymal stem cells and an application thereof. Background technique [0002] At present, the expansion method for obtaining a large number of mesenchymal stem cells at home and abroad is still to add fetal bovine serum (FBS). Although fetal bovine serum contains a variety of nutrients required for cell proliferation, it is widely used in culturing mesenchymal stem cells. Stem cells, however, mesenchymal stem cells cultured in FBS often have problems such as excessive cell aging, many miscellaneous cells, and easy differentiation, and FBS contains a variety of unknown animal-derived proteins, and may even carry bacteria, viruses, and infectious protein diseases Wait. [0003] With the gradual emergence of various unfavorable factors of serum culture, the serum culture program is increasingly rejected by the industry. With the deepening of mesench...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775
CPCC12N5/0668C12N2501/15C12N2500/38C12N2500/90C12N2500/36C12N2500/25C12N2501/998C12N2501/115C12N2501/11C12N2501/12C12N2501/165C12N2501/135C12N2501/105C12N2501/10C12N2500/44C12N2500/46C12N2500/22C12N2500/33C12N2501/392C12N2513/00Y02A50/30
Inventor 李天晴冯春
Owner YUNNAN KEY LAB OF PRIMATE BIOMEDICINE RES
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