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Culture medium and method for quickly inducing differentiation of human mesenchymal stem cells to obtain fat

A technology of induction medium and bone marrow mesenchyme, which is applied in the field of stem cell induction, can solve the problems of decreased cell activity and adhesion force, long experiment time, and long induction time, so as to speed up the process of induction and differentiation and shorten the time spent on experiments , the effect of increasing the rate of induced differentiation

Pending Publication Date: 2019-04-30
FIRST HOSPITAL OF SHANXI MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the existing medium for inducing human bone marrow mesenchymal stem cells to differentiate into fat is not only expensive, but also takes a long time to induce and experiment, and it is easy to cause adverse effects such as cell contamination, cell viability and adhesion force decrease

Method used

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  • Culture medium and method for quickly inducing differentiation of human mesenchymal stem cells to obtain fat
  • Culture medium and method for quickly inducing differentiation of human mesenchymal stem cells to obtain fat
  • Culture medium and method for quickly inducing differentiation of human mesenchymal stem cells to obtain fat

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Embodiment 1

[0035] This embodiment provides a medium for rapidly inducing human bone marrow mesenchymal stem cells to differentiate into fat and an induction method thereof. The specific method includes the following steps:

[0036] (1) FBS and penicillin and streptomycin with a volume ratio of 10:1:86 are added to the IMDM medium and mixed to obtain a complete medium;

[0037] (2) Add dexamethasone to the complete medium, and the concentration of dexamethasone after mixing is 0.8 μM to obtain medium 1;

[0038] (3) Dilute insulin 33 times with pure water to obtain diluted insulin;

[0039] (4) adding diluted insulin to the culture medium 1 described in step (2), the concentration of the mixed insulin is 8 ng / ml to obtain the culture medium 2;

[0040] (5) Mix after adding 3-isobutyl-1-methylxanthine and rosiglitazone in medium 2, the concentration of gained 3-isobutyl-1-methylxanthine is 0.3mM, Rogge The concentration of Litazone is 0.8 μM, then DMSO with a volume fraction of 1:498 is ...

Embodiment 2

[0047] This embodiment provides a medium for rapidly inducing human bone marrow mesenchymal stem cells to differentiate into fat and an induction method thereof. The specific method includes the following steps:

[0048] (1) FBS and penicillin and streptomycin with a volume ratio of 10:1:92 are added to the IMDM medium and mixed to obtain a complete medium;

[0049] (2) Add dexamethasone to the complete medium, and the concentration of dexamethasone after mixing is 1.2 μM to obtain medium 1;

[0050](3) Dilute insulin 37 times with pure water to obtain diluted insulin;

[0051] (4) adding diluted insulin to the culture medium 1 described in step (2), the concentration of the mixed insulin is 12ng / ml to obtain the culture medium 2;

[0052] (5) Mix after adding 3-isobutyl-1-methylxanthine and rosiglitazone in medium 2, the concentration of gained 3-isobutyl-1-methylxanthine is 0.7mM, Rogge The concentration of Litazone is 1.2 μM, then DMSO with a volume fraction of 1:502 is a...

Embodiment 3

[0059] This embodiment provides a medium for rapidly inducing human bone marrow mesenchymal stem cells to differentiate into fat and an induction method thereof. The specific method includes the following steps:

[0060] (1) FBS and penicillin and streptomycin with a volume ratio of 10:1:90 are added to the IMDM medium and mixed to obtain a complete medium;

[0061] (2) Add dexamethasone to the complete medium, and the concentration of dexamethasone after mixing is 1 μM to obtain medium 1;

[0062] (3) Dilute insulin 35 times with pure water to obtain diluted insulin;

[0063] (4) adding diluted insulin to the culture medium 1 described in step (2), the concentration of the mixed insulin is 10 ng / ml to obtain the culture medium 2;

[0064] (5) Mix after adding 3-isobutyl-1-methylxanthine and rosiglitazone in medium 2, the concentration of gained 3-isobutyl-1-methylxanthine is 0.5mM, Luo The concentration of the glitazone is 1 μM, then DMSO with a volume fraction of 1:500 is ...

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Abstract

The invention relates to a culture medium and method for quickly inducing the differentiation of human mesenchymal stem cells to obtain fat. According to the method, at first, an IMDM culture medium is improved: FBS, penicillin, streptomycin, dexamethasone, diluted insulin, 3-isobutyl-1-methylxanthine and rosiglitazone are added into the IMDM culture medium in sequence to obtain an inducing culture medium; primary human mesenchymal stem cells are subjected to subculture; then the inducing culture medium is used to culture human mesenchymal stem cells, and finally the human mesenchymal stem cells are differentiated to obtain fat cells. In the inducing culture medium, dexamethasone, insulin, 3-isobutyl-1-methylxanthine and rosiglitazone all have a fat differentiating inducing function. The provided culture medium and method can accelerate the induced differentiation, shorten the experiment time, increase the induced differentiation rate at the same time, reduce the chance that cells arepolluted, and guarantee the activity of cells.

Description

technical field [0001] The invention belongs to the field of stem cell induction, and in particular relates to a culture medium for rapidly inducing human bone marrow mesenchymal stem cells to differentiate into fat and an induction method thereof. Background technique [0002] Stem cells are a general term for undifferentiated cells before differentiation that can be obtained from various tissues. With the development of economy and society, stem cell research has become a research hotspot, especially in translational medicine in recent years. The biological functions of stem cells, such as proliferation and differentiation, are the hotspots of basic research on stem cells, and they are also an identity mark of stem cells. Human bone marrow mesenchymal stem cells have the characteristics of easy acquisition, fast proliferation, and multi-directional differentiation ability, so human bone marrow mesenchymal stem cells are also mostly used in research. [0003] As one of th...

Claims

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Application Information

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IPC IPC(8): C12N5/077
CPCC12N5/0653C12N2500/30C12N2502/1358C12N2501/33
Inventor 何生姜增誉陈丹阎长平李健丁
Owner FIRST HOSPITAL OF SHANXI MEDICAL UNIV
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