Biological preparation for skin damage repair and preparation method thereof

A biological preparation, a technology for skin damage, which is applied in the field of biological preparations for skin damage repair and their preparation, can solve the problems that the curative effect needs to be improved, the effect is slow, and the treatment course is long.

Inactive Publication Date: 2018-11-23
GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the slow effect of traditional Chinese medicine, the treatment course is longer, and the curative effect needs to be improved

Method used

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  • Biological preparation for skin damage repair and preparation method thereof
  • Biological preparation for skin damage repair and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Embodiment 1 The preparation of biological agent of the present invention

[0030] 1. Take frozen P3 adipose-derived mesenchymal stem cells, thaw the cells in 37°C water, then add the cells to a 15ml centrifuge tube containing 5ml DMEM / F12 medium, centrifuge at 800rpm for 500min, add 5ml DMEM / F12 medium Resuspended, transferred to a 15 cm dish containing DMEM high glucose complete medium containing 10% FBS, in 5% CO 2 , 37°C, saturated humidity of 95% CO 2 cultured in an incubator. When the confluence of mesenchymal stem cells reached 80%, the supernatant was collected.

[0031] 2. Wash the above-mentioned mesenchymal stem cells twice with PBS cleaning solution, take the culture dish, add 20ml of DMEM / F12 basal medium containing 50μg / ml β-glucan, and transfer the washed mesenchymal stem cells to the culture dish at 5% CO 2 , 37°C, saturated humidity of 95% CO 2 Cultivate in an incubator and collect the supernatant;

[0032] 3. Take the cell culture after collectin...

Embodiment 2

[0034] Embodiment 2 The preparation of biological agent of the present invention

[0035] 1. Take frozen P3 adipose-derived mesenchymal stem cells, thaw the cells in 37°C water, then add the cells to a 15ml centrifuge tube containing 5ml DMEM / F12 medium, centrifuge at 800rpm for 500min, add 8ml DMEM / F12 medium Resuspended, transferred to a 15 cm dish containing DMEM high glucose complete medium containing 10% FBS, in 5% CO 2 , 37°C, saturated humidity of 95% CO 2 cultured in an incubator. When the confluence of the mesenchymal stem cells reached 90%, the supernatant was collected.

[0036] 2. Wash the above-mentioned mesenchymal stem cells twice with cleaning solution, take the culture dish, add 20ml of DMEM / F12 basal medium containing 50μg / ml β-glucan, and transfer the washed mesenchymal stem cells to the culture dish , at 5% CO 2 , 37°C, saturated humidity of 95% CO 2 Cultivate in an incubator and collect the supernatant;

[0037] 3. Take the cell culture after collect...

Embodiment 3

[0039] Embodiment 3 detects the proliferation of human skin fibroblasts

[0040] MTT colorimetry was used to detect the effect of the biological preparation prepared in Example 1 of the present invention on the proliferation of human skin fibroblast cell line (HSAS1, P1 generation, purchased from Shanghai Hongshun Biotechnology Co., Ltd.).

[0041] The detection process of this experiment adopts culture medium A, B, C, wherein, culture medium A is the biological preparation that the embodiment 1 of the present invention makes; The obtained conditioned medium was collected; medium C was obtained from the P3 generation bone marrow mesenchymal stem cells and collected according to the culture method in Example 1.

[0042] The determination method is as follows:

[0043] (1) HSAS1 cells were digested and centrifuged to make a cell suspension, diluted with culture medium to a density of 1.5×107L-1, and inoculated into a 96-well culture plate, 200 μL per well.

[0044] (2) After cul...

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Abstract

The invention relates to the technical field of biology, in particular to a preparation method of a biological preparation for skin damage repair. The preparation method comprises the steps that P3 substituted adipose derived stem cells are taken to be recovered and added into a DMEM (dulbecco's modified eagle medium) high glucose complete medium containing 10-12% of FBS (fetal bovine serum) to becultured till the cell confluence rate is as high as 80-90%, and supernatant is collected; the P3 substituted adipose derived stem cells are washed and added into a DMEM / F12 basal culture medium to be cultured for 24 hours, and supernatant is collected; the cell culture after the supernatant is collected is taken, the culture medium is replaced, culture is conducted continuously, the culture liquid is collected, and the steps are repeated for 1-2 times; the supernatants are combined, and the biological preparation is obtained. The biological preparation prepared by adopting the preparation method provided by the invention can outstandingly promote skin fibroblast proliferation and collagen secretion, and has the good effect of promoting skin damage repair.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a biological preparation for repairing skin damage and a preparation method thereof. Background technique [0002] Adipose-derived stem cells (ADSCs) are a kind of stem cells isolated from adipose tissue in recent years with multipotential differentiation potential. Studies have found that ADSCs cells can proliferate stably in vitro and have a low death rate. At the same time, it has the advantages of easy material collection, a large number of stem cells can be obtained from a small amount of tissue, suitable for large-scale culture, and less damage to the body. Moreover, it has a wide range of sources and a large reserve in the body. Suitability for autologous transplantation has gradually become one of the new research hotspots in recent years. At present, adipose stem cells are mainly used in cosmetic industries such as breast enhancement and wrinkle removal in adipose tissue tr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775A61K35/28A61P17/02A61K8/99A61Q19/00A23L33/00
CPCA23L33/00A23V2002/00A61K8/99A61K35/28A61P17/02A61Q19/00C12N5/0667C12N2500/34C12N2500/84A23V2200/318
Inventor 陈海佳葛啸虎王一飞张文祺王小燕
Owner GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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