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Application of insulin-like growth factor 2 in promoting the differentiation of human skin fibroblasts into adipocytes

A fibroblast and insulin-like technology, applied in animal cells, vertebrate cells, cell culture active agents, etc., can solve the problem of low efficiency of skin fibroblast differentiation into adipocytes, difficult wound repair, and scar repair lipotrophy Adverse disease treatment and other issues, to achieve the effect of efficiently inducing differentiation and improving efficiency

Active Publication Date: 2021-12-10
PEKING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] CN201910876049.6 discloses a method for inducing human skin fibroblasts to differentiate into adipocytes in vitro. Using this method, the efficiency of human skin fibroblasts to differentiate into adipocytes is relatively low, and it is difficult to be widely used clinically for wound repair, scarring, etc. Restoration and treatment of lipodystrophy, etc.

Method used

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  • Application of insulin-like growth factor 2 in promoting the differentiation of human skin fibroblasts into adipocytes
  • Application of insulin-like growth factor 2 in promoting the differentiation of human skin fibroblasts into adipocytes
  • Application of insulin-like growth factor 2 in promoting the differentiation of human skin fibroblasts into adipocytes

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0077] Embodiment 1 Efficient in vitro method for inducing human skin fibroblasts to differentiate into adipocytes

[0078] In this example, 24-hour pretreatment with 50 ng / ml insulin-like growth factor 2 (IGF2) was taken as an example to illustrate the results of differentiation induction.

[0079] The method of inducing differentiation is as follows:

[0080] 1. Passage the isolated primary HDF cells to a 35mm culture dish covered with a cover glass, add 3ml of basal medium, and place in a place containing 5% CO 2 in a cell culture incubator at 37°C;

[0081] 2. After the cell confluency reaches 50-70%, add insulin-like growth factor 2 with a final concentration of 50ng / ml, and pretreat the cells for 24 hours;

[0082] 3. Suck off the basal medium, add the induction differentiation medium ①, and continue the induction culture for 6 days, and replace the fresh medium every two days;

[0083] 4. Suck off the differentiation induction medium ①, add the differentiation induct...

Embodiment 2

[0092] Example 2 Efficiently Inducing Human Skin Fibroblasts to Differentiate into Adipocytes in Vitro

[0093] This example takes 100 ng / ml insulin-like growth factor 2 (IGF2) pretreatment for 24 hours as an example to illustrate the results of differentiation induction.

[0094] The method of inducing differentiation is the same as in Example 1, and the "6+2+8" mode of inducing differentiation is also adopted. The difference is that the pretreatment was changed to 100ng / ml IGF-2 pretreatment for 24 hours.

[0095] After the end of the induced differentiation, the results of the induced differentiation were detected by oil red staining and detection of mature adipocyte marker genes. The result is as follows:

[0096] ①Oil red staining results

[0097] Such as Figure 4 As shown, the red part is the stained fat, and the uninduced HDF cells have no fat accumulation, while the HDF cells induced to differentiate for 16 days have obvious fat accumulation. And compared to the ...

Embodiment 3

[0102] Example 3 Efficiently Inducing Human Skin Fibroblasts to Differentiate into Adipocytes in Vitro

[0103] This example takes 200ng / ml insulin-like growth factor 2 (IGF2) pretreatment for 24 hours as an example to illustrate the results of differentiation induction.

[0104] The method of inducing differentiation is the same as in Example 1, and the "6+2+8" mode of inducing differentiation is also adopted. The difference is that the pretreatment was changed to 200ng / ml IGF-2 pretreatment for 24 hours.

[0105] After the end of the induced differentiation, the results of the induced differentiation were detected by oil red staining and detection of mature adipocyte marker genes. The result is as follows:

[0106] ①Oil red staining results

[0107] Such as Figure 7 As shown, the red part is the stained fat, and the uninduced HDF cells have no fat accumulation, while the HDF cells induced to differentiate for 16 days have obvious fat accumulation. And compared with the...

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Abstract

The invention provides the application of insulin-like growth factor 2 in promoting the differentiation of human skin fibroblasts into adipocytes, which is to use insulin-like growth factor 2 with a final concentration of 50-200ng / ml before human skin fibroblasts are induced to differentiate in vitro. Cells were pretreated with growth factor 2. Based on the original "6+2+8" induction differentiation mode, the present invention significantly improves the differentiation efficiency of human skin fibroblasts into fat cells through pretreatment of insulin-like growth factor 2. The method can efficiently induce the differentiation of human skin fibroblasts into adipocytes, and provides new ideas and methods for clinical skin wound healing, scar repair and treatment of lipodystrophy.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the application of insulin-like growth factor 2 in promoting the differentiation of human skin fibroblasts into fat cells. Background technique [0002] CN201910876049.6 discloses a method for inducing human skin fibroblasts to differentiate into adipocytes in vitro. Using this method, the efficiency of human skin fibroblasts to differentiate into adipocytes is relatively low, and it is difficult to be widely used clinically for wound repair, scarring, etc. Restoration and treatment of lipodystrophy, etc. Contents of the invention [0003] The purpose of the present invention is to provide the application of insulin-like growth factor 2 in promoting the differentiation of human skin fibroblasts into adipocytes. [0004] Another object of the present invention is to provide a method for efficiently inducing human skin fibroblasts to differentiate into adipocytes in vitro. [0005]...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/077
CPCC12N5/0653C12N2501/105C12N2501/33C12N2500/02C12N2523/00C12N2506/1307
Inventor 张传茂王向阳
Owner PEKING UNIV
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