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Application of vitamin D3 and analogues thereof in promoting differentiation of human skin fibroblasts into adipocytes

A fibroblast and adipocyte technology, applied in the biological field, can solve the problems of difficult wound repair, scar repair lipodystrophy treatment, low efficiency of skin fibroblast differentiation into adipocytes, etc., and achieve efficient induction of differentiation into , the effect of improving efficiency

Active Publication Date: 2020-08-11
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 vitamin D3 and analogues thereof in promoting differentiation of human skin fibroblasts into adipocytes
  • Application of vitamin D3 and analogues thereof in promoting differentiation of human skin fibroblasts into adipocytes
  • Application of vitamin D3 and analogues thereof in promoting differentiation of human skin fibroblasts into adipocytes

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0081] Embodiment 1 Efficient in vitro method for inducing human skin fibroblasts to differentiate into adipocytes

[0082] In this example, 24-hour pretreatment with 10 nM vitamin D3 (Calcitriol) was taken as an example to illustrate the results of differentiation induction.

[0083] The method of inducing differentiation is as follows:

[0084] 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;

[0085] 2. After the cell confluency reaches 50-70%, add vitamin D3 with a final concentration of 10nM, and pretreat the cells for 24 hours;

[0086] 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;

[0087] 4. Suck off the differentiation induction medium ①, add the differentiation induction medium ②, and induce culture for...

Embodiment 2

[0096] Example 2 Efficiently Inducing Human Skin Fibroblasts to Differentiate into Adipocytes in Vitro

[0097] In this example, 50 nM vitamin D3 (Calcitriol) was pretreated for 24 hours as an example to illustrate the results of differentiation induction.

[0098] 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 50 nM vitamin D3 pretreatment for 24 hours.

[0099] 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:

[0100] ①Oil red staining results

[0101] 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 with the control grou...

Embodiment 3

[0106] Example 3 Efficiently Inducing Human Skin Fibroblasts to Differentiate into Adipocytes in Vitro

[0107] In this example, 10 nM calcipotriol (Calcipotriol) was pretreated for 24 hours as an example to illustrate the results of differentiation induction.

[0108] 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 10 nM calcipotriol pretreatment for 24 hours.

[0109] 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:

[0110] ①Oil red staining results

[0111] 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 to the control ...

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Abstract

The invention provides an application of vitamin D3 and analogues thereof in promoting differentiation of human skin fibroblasts into adipocytes. The application is characterized in that cells are pretreated by using vitamin D3 or analogues thereof with the final concentration of 10-100nM before the human skin fibroblasts are induced and differentiated in vitro. On the basis of an original 6 + 2 +8 induced differentiation mode, the efficiency of differentiating human skin fibroblasts into adipocytes is remarkably improved through pretreatment of vitamin D3 or the analogues thereof. By adopting the method, human skin fibroblasts can be efficiently induced and differentiated into adipocytes, and a new idea and a new method are provided 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 vitamin D3 and its analogues 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 vitamin D3 and its analogs in promoting the differentiation of human skin fibroblasts into fat cells. [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] In...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/077
CPCC12N5/0653C12N2506/1307C12N2500/30C12N2500/38C12N2501/33
Inventor 张传茂王向阳
Owner PEKING UNIV
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