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A method for improving the adipogenic differentiation efficiency of bone marrow mesenchymal stem cells

A bone marrow mesenchymal and adipogenic differentiation technology, applied in the field of stem cell differentiation and biomedicine, can solve problems such as low efficiency of adipogenic differentiation, achieve high repeatability, meet clinical application requirements, and have no cytotoxic effects.

Active Publication Date: 2019-08-23
SHANGHAI NAT ENG RES CENT FORNANOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] The object of the present invention is to aim at the inefficiency of adipogenic differentiation of MSCs on the surface of commercialized cell culture substrates, and provide a method that can greatly increase the ratio of adipogenic differentiation of MSCs by changing the replacement mode of the cell culture medium

Method used

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  • A method for improving the adipogenic differentiation efficiency of bone marrow mesenchymal stem cells
  • A method for improving the adipogenic differentiation efficiency of bone marrow mesenchymal stem cells
  • A method for improving the adipogenic differentiation efficiency of bone marrow mesenchymal stem cells

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Embodiment 1

[0026] Rat bone marrow mesenchymal stem cells were selected as the cell model, and MSCs were extracted from the bone marrow cavity of tibia and femur of newborn SD rats by whole bone marrow adherent method and cultured. The culture medium of MSC was low-sugar DMEM medium containing 10% (volume ratio) fetal bovine serum (FBS), 100 U / mL penicillin, 100 μg / mL streptomycin and 2 mM L-glutamine. After MSC extraction, store at 37 °C, 5% CO 2 The primary culture was carried out in an incubator, and the new culture medium was replaced every 2 to 3 days until the MSCs basically covered the bottom of the culture bottle, and then subcultured. The passage time does not need to be too early.

[0027] When performing cell subculture, first perform digestion and centrifugation. After the centrifugation, suck out the supernatant, that is, the original cell culture medium, and do not discard it, but mix it with the newly prepared cell culture medium at a ratio of 80:20. Volume ratio was mixe...

Embodiment 2

[0030] Rat bone marrow mesenchymal stem cells were selected as the cell model, and MSCs were extracted from the bone marrow cavity of tibia and femur of newborn SD rats by whole bone marrow adherent method and cultured. The culture medium of MSC was low-sugar DMEM medium containing 10% (volume ratio) fetal bovine serum (FBS), 100 U / mL penicillin, 100 μg / mL streptomycin and 2 mM L-glutamine. After MSC extraction, store at 37 °C, 5% CO 2 The primary culture was carried out in an incubator, and the new culture medium was replaced every 2 to 3 days until the MSCs basically covered the bottom of the culture bottle, and then subcultured. The passage time does not need to be too early.

[0031] When performing cell subculture, first perform digestion and centrifugation. After the centrifugation, suck out the supernatant, that is, the original cell culture medium. Instead of discarding it, mix it with the newly prepared cell culture medium at a ratio of 50:50. Mix the volume ratio, ...

Embodiment 3

[0034] Rat bone marrow mesenchymal stem cells were selected as the cell model, and MSCs were extracted from the bone marrow cavity of tibia and femur of newborn SD rats by whole bone marrow adherent method and cultured. The culture medium of MSC is low-glucose DMEM medium containing 10% (volume ratio) fetal bovine serum (FBS), 100 U / mL penicillin, 100 μg / mL streptomycin and 2 mM L-glutamine. After MSC extraction, store at 37 °C, 5% CO 2 The primary culture was carried out in an incubator, and the new culture medium was replaced every 2 to 3 days until the MSCs basically covered the bottom of the culture bottle, and then subcultured. The passage time does not need to be too early.

[0035] When performing cell subculture, first perform digestion and centrifugation. After centrifugation, suck out the supernatant, that is, the original cell culture medium, and do not discard it, but mix it with the newly prepared cell culture medium at a ratio of 20:80. The volume ratio was mix...

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Abstract

The invention relates to a method for improving the adipogenic differentiation efficiency of mesenchymal stem cells. The method comprises the following steps: performing extraction and primary culture on MSC, performing subculture on MSC, and performing adipogenic differentiation characterization on MSC. According to the method, adipose cells of different quantities are obtained by controlling the replacement quantity of a culture solution and a cell culture period during subculture, so that extremely high MSC adipogenic differentiation efficiency can be still obtained on the surface of a polystyrene cell culture substrate under the condition that an inducible factor is not contained. The method has the characteristics of simple system, absence of extra biological factors, cytotoxicity avoidance, high repeatability and the like. The method is capable of meeting the in-vitro stem cell culture requirements and clinical application requirements.

Description

technical field [0001] The invention relates to a method for improving the adipogenic differentiation efficiency of bone marrow mesenchymal stem cells, in particular to a method for improving the bone marrow on the surface of the commercialized cell culture medium by changing the replacement mode of the cell culture medium without adding an inducer. Methods for Adipogenic Differentiation Efficiency of Mesenchymal Stem Cells. The invention belongs to the field of stem cell differentiation and biomedicine. Background technique [0002] Stem cells are a type of cells with strong self-renewal ability and multi-directional differentiation potential, and can differentiate into different tissue cell types under certain conditions. Therefore, stem cells have become an ideal seed cell type in tissue repair and play a pivotal role in tissue engineering research ( Nature , 2009, 462: 433). According to the source of origin, stem cells can be divided into embryonic stem cells and adu...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/077
CPCC12N5/0653C12N2506/1353C12N2533/30
Inventor 何丹农叶恺王萍金彩虹
Owner SHANGHAI NAT ENG RES CENT FORNANOTECH
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