Preparation method and product of fibroblasts for skin scar repair

A technology of fibroblasts and fibroblasts, applied in the field of preparation of fibroblasts for skin scar repair and its products, can solve problems such as pigmentation and complications, and inability to remove scars

Pending Publication Date: 2018-11-13
岳萍
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, surgical resection is the main method for relatively serious depressed scars, atrophic scars, and hypertrophic scars, but any surgical method cannot completely remove the scars, but can only improve or correct the harm caused by the scars to the greatest extent. new scars
For patients with milder scars, laser, cryotherapy or radiotherapy are more commonly used methods, and the effect is better, but pigmentation and complications are prone to occur

Method used

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  • Preparation method and product of fibroblasts for skin scar repair
  • Preparation method and product of fibroblasts for skin scar repair
  • Preparation method and product of fibroblasts for skin scar repair

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] The preparation of embodiment 1 human fibroblast

[0048] The discarded skin was washed three times with 1×PBS (pH 7.4) containing 2 times penicillin and streptomycin, and then 5-20 mL of 0.1% type II collagenase (purchased by Sigma, USA) at a mass volume ratio of 1:1 was used. Digest overnight in a refrigerator at 4°C; after digestion, take dermis tissue with a mass volume ratio of 0.25% trypsin (containing 0.02% EDTA in mass volume ratio) (purchased by Thermo Fisher, USA) and digest it in a 37°C incubator for 4 hours. Then centrifuge at 1500rpm for 10 minutes, discard the supernatant, add normal saline to resuspend, and filter with 100μm cell sieve; cell suspension is centrifuged at 1000rpm for 10 minutes, cell pellet is resuspended and washed twice with normal saline, then add fibroblast serum-free culture medium (Containing 1ng-100ng / ml fibroblast growth factor, purchased by American R&D company; 0.5ng-100ng / ml vitamin E, purchased by American Sigma company; Identi...

Embodiment 2

[0052] The kit product of the fibroblast used for skin scar repair that embodiment 2 prepares

[0053] The kit of fibroblasts for skin scar repair includes:

[0054] 1) 500ml serum-free medium for fibroblasts;

[0055] 2) 10 4 IU / mL type II collagenase;

[0056] 3) 100ml 0.25% trypsin;

[0057] 4) 10mL 50 times active composition: 50ng-5μg fibroblast growth factor, 25ng-5μg vitamin E, 5mg-100mg phosphate-3-O-rutinoside, 5mg-100mg L-lactic acid;

[0058] 5) 10mL 50 times serum-free supplement;

[0059] 6) instruction manual;

[0060] Wherein, the instructions for use include the methods described in Examples 1-3.

Embodiment 3

[0061] Example 3 In vitro co-culture of human fibroblasts inhibits the proliferation of hypertrophic scar fibroblasts

[0062] Hypertrophic scar fibroblasts in the logarithmic growth phase (purchased from ScienCell, USA) were taken in 3 × 10 5 The concentration of cells / well was inoculated in 6-well Transwell culture plates (0.3 μm, Corning Company, USA), cultured in UltraCULTURETM without serum, 1.5 mL was inoculated in each well, and human fibroblasts were added to the small chamber, 3×105 cells / chamber, 1.5 ml, placed at 37 ℃, 5% CO2 incubator co-cultivation, for grouping. Blank control group A: No fibroblast co-culture (Control) was added to the small chamber, 6 repeated wells; fibroblast experiment group B prepared in Example 1: normal fibroblasts (FB) were added to the small chamber for co-culture, repeated wells 6; and the fibroblast control group C of comparison ratio culture (comparison ratio fibroblast adopts " in adding fibroblast growth factor and not adding in th...

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Abstract

The invention provides a preparation method of fibroblasts for skin scar repair, wherein the method comprises the steps of extracting fibroblasts from tissues and culturing with a fibroblast growth factor, vitamin E, phosphoric acid-3-O-rutinoside and L-lactic acid to obtain the fibroblasts. The prepared fibroblasts have good biological activity, repair broken fibrous tissues, restore a collagen structure of a skin basal layer, and also inhibit fibrosis and scar formation caused by overexpression of collagen; and the fibroblasts provided by the invention have good therapeutic and cosmetic effects on skin scars.

Description

technical field [0001] The present invention relates to a method of fibroblasts for repairing skin scars and obtaining fibroblasts for treating skin scars by said method, in particular, the present invention relates to obtaining fibroblasts from tissues through in vitro culture, capable of repairing fractures Fibrous tissue can restore the collagen structure of the basal layer of the skin; at the same time, it can inhibit the fibrosis and scar formation caused by the overexpression of collagen, and it is expected to achieve better therapeutic and cosmetic effects on skin scars. Background technique [0002] Skin scar is the appearance, morphology and histopathological changes of normal skin tissue caused by various traumas, and the fibrous connective tissue formed by the remodeling and maturation of granulation tissue. Excessive proliferation of body tissue, mild hyperplasia of connective tissue, or scar formation caused by mild hyperplasia of connective tissue after skin in...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071A61K35/33A61P17/02
CPCA61P17/02A61K35/33C12N5/0656C12N2500/30C12N2500/38C12N2501/115C12N2500/90C12N2509/00
Inventor 不公告发明人
Owner 岳萍
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