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Exosome secretion culture medium and culture and separation method of umbilical cord mesenchymal stem cell exosomes

A technology of stem cells and culture methods, which is applied in the field of exosome secretion medium and the culture and separation of umbilical cord mesenchymal stem cell exosomes, which can solve the problems of difficult operation in the separation and purification of exosomes.

Active Publication Date: 2021-06-08
广州研华生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Moreover, the small number of exosomes also brings great operational difficulties to downstream separation and purification.

Method used

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  • Exosome secretion culture medium and culture and separation method of umbilical cord mesenchymal stem cell exosomes
  • Exosome secretion culture medium and culture and separation method of umbilical cord mesenchymal stem cell exosomes
  • Exosome secretion culture medium and culture and separation method of umbilical cord mesenchymal stem cell exosomes

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Experimental program
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Effect test

Embodiment 1

[0061] This embodiment provides an exosome secretion medium.

[0062] The mesenchymal serum-free stem cell medium was used as the basal medium, and the following components were added: 200ng / ml EGF, 150ng / ml TGF, 30μg / ml coenzyme Q10, 600μg / ml potassium salt, 0.6μg / ml Fructose diphosphate sodium and 300 μg / ml histamine dihydrochloride.

[0063] The content of each of the above components is the concentration in the mesenchymal serum-free stem cell culture medium.

[0064] Potassium salt is: potassium chloride dissolved in a mixed salt solution of potassium dihydrogen phosphate buffer and dipotassium hydrogen phosphate buffer, the total concentration of potassium dihydrogen phosphate buffer and dipotassium hydrogen phosphate buffer is 0.01mol / L, pH is 7.4, and the concentration of potassium chloride in the mixed salt solution is 1g / L.

Embodiment 2

[0066] This embodiment provides an exosome secretion medium.

[0067] Based on the mesenchymal serum-free stem cell medium, add the following components: EGF: 200ng / ml; TGF: 150ng / m; Q10: 30μg / ml; Potassium salt: 300μg / ml; FDP: 0.6μg / ml ; Histamine dihydrochloride: 300 μg / ml. Potassium salt is: potassium chloride dissolved in a mixed salt solution of potassium dihydrogen phosphate buffer and dipotassium hydrogen phosphate buffer, the total concentration of potassium dihydrogen phosphate buffer and dipotassium hydrogen phosphate buffer is 0.01mol / L, pH is 7.4, and the concentration of potassium chloride in the mixed salt solution is 1g / L.

Embodiment 3

[0069] This embodiment provides an exosome secretion medium.

[0070] The mesenchymal serum-free stem cell medium was used as the basal medium, and the following components were added: 200ng / ml EGF, 150ng / ml TGF, 30μg / ml coenzyme Q10, 600μg / ml potassium salt, 0.3μg / ml Fructose diphosphate sodium and 300 μg / ml histamine dihydrochloride. Potassium salt is: potassium chloride dissolved in a mixed salt solution of potassium dihydrogen phosphate buffer and dipotassium hydrogen phosphate buffer, the total concentration of potassium dihydrogen phosphate buffer and dipotassium hydrogen phosphate buffer is 0.01mol / L, pH is 7.4, and the concentration of potassium chloride in the mixed salt solution is 1g / L.

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Abstract

The invention discloses an exosome secretion culture medium and a culture and separation method of umbilical cord mesenchymal stem cell exosomes, and relates to the technical field of exosome enrichment and separation. A serum-free stem cell culture medium is used as a basic culture medium; and the following components are added: EGF with the concentration of 100 to 300ng / ml, TGF with the concentration of 50 to 150ng / ml, coenzyme Q10 with the concentration of 10 to 30mu g / ml, potassium salt with the concentration of 300 to 600mu g / ml, fructose sodium diphosphate with the concentration of 0.3 to 0.9 mu g / ml and histamine dihydrochloride with the concentration of 100 to 300 mu g / ml. By adopting the exosome secretion culture medium provided by the invention, more exosomes can be obtained under the condition of the same number of culture cells. The exosome secretion culture medium is beneficial to large-scale production of umbilical cord mesenchymal stem cell exosomes with good biological activity, further promotes wide application of the exosomes in medical treatment, anti-aging, beauty treatment and the like, and has good economic benefits.

Description

technical field [0001] The present invention relates to the technical field of exosome enrichment and separation, in particular to an exosome secretion medium and a method for culturing and separating umbilical cord mesenchymal stem cell exosomes. Background technique [0002] Exosomes (exosomes) are a type of membranous vesicles that can be secreted by a variety of cells and encapsulate proteins, mRNA, microRNA (microRNA), non-coding RNA (lncRNA), DNA, lipids and other substances. The body plays an important role in intercellular communication, tumor immunity, tumor therapy, tissue damage repair, and disease diagnosis. American scientists James E. Rothman and Randy W. Schekman, and German scientist Thomas C. Südhof shared the 2013 Nobel Prize in Physiology or Medicine for their discoveries based on the transport regulation mechanism of vesicles (exosomes, etc.) inside cells. [0003] Mesenchymal stem cells (MSCs) are pluripotent stem cells derived from the mesoderm, which ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775
CPCC12N5/0665C12N5/0668C12N2500/90C12N2501/11C12N2501/148C12N2501/15C12N2501/999C12N2500/12C12N2500/42C12N2500/46C12N2501/998C12N2509/00
Inventor 麦志国刘元旦陈群珍尹丽珍
Owner 广州研华生物科技有限公司
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