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Promoting fibroblast transdifferentiation into adipocytes using defined cytokine combinations

A fibroblast and hepatocyte growth factor technology, applied in the direction of animal cells, vertebrate cells, cell culture active agents, etc., can solve the problem of inability to separate adipocytes, and achieve the effect of simple induction process

Inactive Publication Date: 2017-04-05
GUANGZHOU INST OF BIOMEDICINE & HEALTH CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the marker genes of preadipocytes have not been determined so far, so it has not been possible to perfectly isolate the true preadipocytes from the body for research

Method used

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  • Promoting fibroblast transdifferentiation into adipocytes using defined cytokine combinations
  • Promoting fibroblast transdifferentiation into adipocytes using defined cytokine combinations
  • Promoting fibroblast transdifferentiation into adipocytes using defined cytokine combinations

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0076] The transdifferentiation of embodiment 1NIH-3T3 cell

[0077] ABCDE group:

[0078] The revived NIH-3T3 cells were subcultured twice with cell growth medium, and then induced to differentiate. Taking one well of a 24-well plate as an example, after the cells grew and converged, the medium was replaced with cell induction medium (0.5ml) , in the induction medium, the final concentration of cytokines is as follows:

[0079] A: epidermal growth factor, final concentration 20ng / ml;

[0080] B: hepatocyte growth factor, final concentration 20ng / ml;

[0081] C: dexamethasone, final concentration 100nM;

[0082] D: Insulin, 5 μg / ml;

[0083] E: Rosiglitazone, the final concentration is 1 μM;

[0084] The induction medium was changed every two days and cultured for two weeks.

[0085] Blank control group:

[0086] The transdifferentiation culture conditions of NIH-3T3 cells were the same as those of ABCDE group, except that no cell induction medium was used, only cell gr...

Embodiment 2

[0099] Example 2 Transdifferentiation of Mouse Tail Fibroblasts

[0100] Isolation and culture of mouse tail cells: Take mouse tail cells of 3 cm outdoors, soak them in 75% ethanol for 30 seconds, and then transfer them to the medium supplemented with 5 times double antibody. Then use surgical scissors to cut the mouse tail into small pieces; add the medium preheated at 37 degrees Celsius, and after 5 days, you can see that there are cells attached to the wall. After 12 days, digest the cells, plate them in a 12-well culture plate, and wait for the cells to congregate .

[0101] ABCDE group:

[0102] Differentiation of mouse tail fibroblasts: The steps of inducing differentiation were exactly the same as those of NIH-3T3 cells in Example 1, cultured for 5 weeks and observed.

[0103] Blank control group: the culture procedure is the same as that of the blank control test in Example 1.

[0104] Induction group: the steps of inducing differentiation were the same as the MDIR ...

Embodiment 3

[0109] The revived NIH-3T3 cells were subcultured twice with cell growth medium, and then induced to differentiate. Taking one well of a 24-well plate as an example, after the cells grew and converged, the medium was replaced with cell induction medium (0.5ml) , in the induction medium, the final concentration of cytokines is as follows:

[0110] A: epidermal growth factor, final concentration 10ng / ml;

[0111] B: hepatocyte growth factor, final concentration 10ng / ml;

[0112] C: dexamethasone, final concentration 100nM;

[0113] D: Insulin, 1 μg / ml;

[0114] E: Rosiglitazone, the final concentration is 0.5 μM;

[0115] The induction medium was changed every two days for four weeks.

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Abstract

Provided in the present invention is a composition for promoting the transdifferentiation of fibroblasts into adipocytes, the onset factors thereof being a hepatocyte growth factor, epidermal growth factor, dexamethasone and insulin. Also provided in the present invention is a culture solution and a method for inducing the transdifferentiation of fibroblasts into adipocytes.

Description

technical field [0001] The invention relates to a cell transdifferentiation-inducing cytokine composition and a method for culturing cell transdifferentiation, in particular to a cytokine composition for promoting the transdifferentiation of fibroblasts into adipocytes and using the composition to promote the transdifferentiation of fibroblasts into fat Cell culture method. Background technique [0002] The cause of obesity is that when the energy in the body is out of balance and the energy intake is greater than the energy consumed, the excess energy is mainly stored in the form of fat, and the weight of fat increases, leading to the occurrence of obesity. Fat is mainly stored in the adipose tissue of the body, and the increase of adipose tissue is mainly manifested in the increase of the volume of fat cells and the increase of the number of fat cells. However, the volume of fat cells has a certain limit, and it is impossible to increase indefinitely, so excessive energy ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/077
CPCC12N5/0653C12N2501/11C12N2501/12C12N2501/33C12N2501/39C12N2506/1307
Inventor 吴东海聂涛徐爱民李鹏
Owner GUANGZHOU INST OF BIOMEDICINE & HEALTH CHINESE ACAD OF SCI
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