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Mesenchymal stem cell adipogenesis induced differentiation method

A mesenchymal stem cell and induced differentiation technology, applied in biochemical equipment and methods, animal cells, vertebrate cells, etc., can solve the problem of increased operational difficulty and cost, no mesenchymal stem cell adipogenic induction differentiation method, differentiation medium Complex components and other issues, to achieve the effect of less reagent components, simplified induction differentiation operation, and proper operation

Inactive Publication Date: 2019-07-09
上海葆年生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0014] However, the composition of the induction differentiation medium used in the adipogenic differentiation process of mesenchymal stem cells is still relatively complicated, which increases the difficulty and cost of operation
There is no simpler and more efficient method for adipogenic induction of mesenchymal stem cells

Method used

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  • Mesenchymal stem cell adipogenesis induced differentiation method
  • Mesenchymal stem cell adipogenesis induced differentiation method
  • Mesenchymal stem cell adipogenesis induced differentiation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] 1. Prepare umbilical cord mesenchymal stem cell adipogenic differentiation medium (homemade DMEM / F12 basic medium, 10% fetal bovine serum, 10μg / ml insulin, 1‰ penicillin, 1‰ streptomycin, 1μmol / L dexamethasone , 300μmol / L 3-isobutyl-1-methylxanthine).

[0041] 2. After the cells are recovered, the cells to be tested will grow to about 80% for passage. Generally do 3 holes in a six-well plate, one uninduced control, one experimental group, and one duplicate hole.

[0042] 3. Digest the mesenchymal stem cells according to 1×10 4 cells / cm 2 Inoculate the cell density in a six-well plate, add 3ml DMEM complete medium to each well, and place the cells at 37℃, 5% CO 2 Culture in the incubator.

[0043] 4. About 2 days, the cell fusion degree reaches about 80%, carefully aspirate the complete medium of mesenchymal stem cells, and add 3ml of mesenchymal stem cell adipogenic differentiation medium to the two experimental groups of the six-well plate.

[0044] 5. Change to fresh mesench...

Embodiment 2

[0047] 1. Prepare umbilical cord mesenchymal stem cell adipogenic differentiation medium (home-made DMEM / F12 basic medium, 8% fetal calf serum, 8μg / ml insulin, 0.8‰ penicillin, 1‰ streptomycin, 1.2μmol / L dexamethasone Methosone, 500μmol / L 3-isobutyl-1-methylxanthine).

[0048] 2. After the cells are resuscitated, the cells to be tested will grow to about 90% for passage. Generally do 3 holes in a six-well plate, one uninduced control, one experimental group, and one duplicate hole.

[0049] 3. Digest the mesenchymal stem cells according to 1×10 4 cells / cm 2 Inoculate the cell density in a six-well plate, add 3ml DMEM complete medium to each well, and place the cells at 37℃, 5% CO 2 Culture in the incubator.

[0050] 4. About 1 day, the cell fusion degree reaches about 70%, carefully aspirate the complete medium of mesenchymal stem cells, and add 3ml of mesenchymal stem cell adipogenic differentiation medium to the two experimental groups of the six-well plate.

[0051] 5. Change to f...

Embodiment 3

[0054] 1. Prepare umbilical cord mesenchymal stem cell adipogenic differentiation medium (home-made DMEM / F12 basic medium, 10% fetal calf serum, 12μg / ml insulin, 0.8‰ penicillin, 1.2‰ streptomycin, 0.8μmol / L dexamethasone Methosone, 100μmol / L 3-isobutyl-1-methylxanthine).

[0055] 2. After the cells are recovered, the cells to be tested will grow to about 80% for passage. Generally do 3 holes in a six-well plate, one uninduced control, one experimental group, and one duplicate hole.

[0056] 3. Digest the mesenchymal stem cells according to 1×10 4 cells / cm 2 Inoculate the cell density in a six-well plate, add 3ml DMED complete medium to each well, and place the cells at 37℃, 5% CO 2 Culture in the incubator.

[0057] 4. About 3 days, the cell fusion degree reached about 80%, carefully aspirate the complete medium of mesenchymal stem cells, and add 3ml of mesenchymal stem cell adipogenic differentiation medium to the two experimental groups of the six-well plate.

[0058] 5. Change to...

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Abstract

The invention discloses a mesenchymal stem cell adipogenesis induced differentiation method. The method includes the following steps: (1) recovering the mesenchymal stem cells to grow to 70-90%; (2) digesting the mesenchymal stem cells and culturing for 1-3 days by using a complete culture medium; and (3) when the cell fusion degree reaches 70-90%, sucking off the complete culture medium, adding amesenchymal stem cell fat-forming induction differentiation culture medium (composed of DMEM / F12 basal culture medium, fetal bovine serum with volume concentration of 8-12%, 8-12 ug / ml of insulin, 1%. of penicillin, 1 %. of streptomycin, 0.8-1.2 umol / L of dexamethasone, 100-500 umol / L of 3-isobutyl-1-methylxanthine) for culture, and changing the culture medium every 3-4 days; preforming continuous induction, which depends on cell growth and adipogenesis condition. The method is proper in operation, simplifies the steps of induced differentiation and is quite high in adipogenesis induced differentiation rate.

Description

Technical field [0001] The present invention relates to the field of biotechnology, in particular to a method for adipogenic induction and differentiation of mesenchymal stem cells. Background technique [0002] Mesenchymal stem cells (MSC) derived from embryonic mesoderm are pluripotent stem cells with high self-renewal ability and multi-differentiation potential. They are widely found in bone marrow, fat, amniotic membrane, placenta, cord blood and umbilical cord tissue It can induce differentiation into adipocytes, osteoblasts, cardiomyocytes, pancreatic islet-like cells, vascular endothelial cells and epidermal cells in vitro. In addition, human mesenchymal stem cells have low immunogenicity, low risk of pathogen infection and tumorigenicity, and high safety in clinical application. In addition, it is simple and convenient to obtain materials without ethical restrictions. After continuous subculture and cryopreservation, its biological characteristics are still maintained. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/077
CPCC12N5/0653C12N2500/40C12N2501/30C12N2501/33C12N2506/1346
Inventor 崔晓燕徐卓
Owner 上海葆年生物科技有限公司
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