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Medium for amplification of human mesenchymal stem cells and amplification method thereof

A mesenchymal stem cell and culture medium technology, applied in the field of mesenchymal stem cell culture, can solve the problems of slow expansion of expansion medium, easy differentiation of stem cells, and long expansion time, so as to inhibit the growth of cancer cells, shorten the The effect of expanding time and enhancing cell activity

Inactive Publication Date: 2018-02-02
金时代进出口贸易重庆有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] Aiming at the above-mentioned deficiencies in the prior art, the present invention provides a culture medium for expanding human mesenchymal stem cells and an expansion method thereof, which can effectively solve the problem of slow expansion speed and time-consuming expansion of existing expansion medium. long, stem cells are easy to differentiate, etc.

Method used

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  • Medium for amplification of human mesenchymal stem cells and amplification method thereof

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Embodiment 1

[0030] A medium for expanding human mesenchymal stem cells, including a basal medium and additives added to the medium, the basal medium is DMEM / F12 medium, and the additives include components of the following final concentrations: linoleic acid 2 μL / mL, human AB type serum 5μL / mL, human transferrin 5μg / mL, L-glutamine 1.5mmol / mL, vitamin C 2mg / mL, grape seed extract 0.5mg / mL, salidroside 1mg / mL, ginsenoside 5mg / mL, Lycium barbarum polysaccharide 2mg / mL, donggui phthalide 0.2mg / mL, platelet-derived growth factor 0.1μL / mL and epidermal growth factor 0.1μL / mL.

[0031] Among them, salidroside is prepared by the following method: mixing rhodiola rosea and water at a solid-to-liquid ratio of 1:20, soaking at room temperature for 0.5h, decocting for 0.5h, filtering, and adding medicinal residues 6 Double the weight of water, continue to decoct for 0.5h, filter, combine the two filtrates, then concentrate to a relative density of 1.1-1.2, finally add ethanol to make the volume pe...

Embodiment 2

[0040]A medium for expanding human mesenchymal stem cells, including a basal medium and additives added to the medium, the basal medium is DMEM / F12 medium, and the additives include the following final concentrations of components: 10 μL of linoleic acid / mL, human AB type serum 15μL / mL, human transferrin 15μg / mL, L-glutamine 3mmol / mL, vitamin C 5mg / mL, grape seed extract 1mg / mL, salidroside 5mg / mL , 10 mg / mL ginsenoside, 5 mg / mL wolfberry polysaccharide, 0.6 mg / mL donggui phthalide, 0.2 μL / mL platelet-derived growth factor and 0.2 μL / mL epidermal growth factor.

[0041] Among them, salidroside is prepared by the following method: mixing Rhodiola rosea and water at a solid-to-liquid ratio of 2:20, soaking at room temperature for 1 hour, decocting for 1 hour, filtering, and adding 8 times the weight of the drug residue to the drug residue water, continue to decoct for 0.5h, filter, combine the filtrate twice, then concentrate to a relative density of 1.1-1.2, finally add ethano...

Embodiment 3

[0048] A medium for expanding human mesenchymal stem cells, including a basal medium and additives added to the medium, the basal medium is DMEM / F12 medium, and the additives include components of the following final concentrations: 8 μL of linoleic acid / mL, human AB type serum 10μL / mL, human transferrin 10μg / mL, L-glutamine 2.2mmol / mL, vitamin C 3mg / mL, grape seed extract 0.8mg / mL, salidroside 3mg / mL, ginsenoside 6mg / mL, Lycium barbarum polysaccharide 3mg / mL, dongangi phthalide 0.3mg / mL, platelet-derived growth factor 0.1μL / mL and epidermal growth factor 0.2μL / mL.

[0049] Among them, salidroside is prepared by the following method: mixing Rhodiola rosea and water at a solid-to-liquid ratio of 2:20, soaking at room temperature for 1 hour, decocting for 1 hour, filtering, and adding 8 times the weight of the drug residue to the drug residue water, continue to decoct for 0.5h, filter, combine the filtrate twice, then concentrate to a relative density of 1.1-1.2, finally add e...

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Abstract

The invention discloses a culture medium for expanding human mesenchymal stem cells, which comprises a basic medium and an additive added to the medium, and the additive includes the following components at final concentrations: linoleic acid 2‑10 μL / mL, human Source AB type serum 5‑15 μL / mL, human transferrin 5‑15 μg / mL, etc. Using the culture medium to expand human mesenchymal stem cells includes the following steps: (1) isolating human mesenchymal stem cells from umbilical cord or placenta, or isolating mononuclear cells from bone marrow blood, umbilical cord blood or placental blood, and placing them in the culture medium (2) Place the cells separated and purified in step (1) into fresh medium for subculture. The invention adopts the culture medium of specific components and the expansion method, which can effectively speed up the cell expansion speed, shorten the expansion time, maintain the uniform shape of the cells after passaging, and reduce their differentiation, thereby ensuring the safety of human mesenchymal stem cells sex.

Description

technical field [0001] The invention belongs to the technical field of culturing mesenchymal stem cells, and in particular relates to a medium for expanding human mesenchymal stem cells and an expanding method thereof. Background technique [0002] Mesenchymal stem cells (MSCs) are an important member of the stem cell family, derived from the mesoderm and ectoderm in the early stages of development, belonging to pluripotent stem cells, which can differentiate into pancreatic islets, neural , vascular endothelium, bone, cartilage, muscle, liver, cardiac muscle and other tissue cells. MSCs have become the seed cells for cell and gene therapy research because of their relative lack of immunogenicity, simple culture technology, and cryopreservation. They have potential clinical application value and have become a hot spot in stem cell research. [0003] At present, the medium used for the cultivation and expansion of mesenchymal stem cells is mainly to add a certain concentrati...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775
CPCC12N5/0665C12N2500/24C12N2500/32C12N2500/36C12N2500/38C12N2500/76C12N2501/11C12N2501/135C12N2501/90C12N2501/999
Inventor 赵翔宋彩霞曹玉昊叶劲涛王仁杰黄启程邱阳李国焱唐玲胡小东
Owner 金时代进出口贸易重庆有限公司
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