Composition comprising mesenchymal stem cells and exosomes and its application in tissue repair

A technology of exosomes and epidermal stem cells, applied in the biological field

Active Publication Date: 2021-09-10
广东肽诺干细胞生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, exosomes have not been used in diabetic tissue repair applications so far.

Method used

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  • Composition comprising mesenchymal stem cells and exosomes and its application in tissue repair
  • Composition comprising mesenchymal stem cells and exosomes and its application in tissue repair
  • Composition comprising mesenchymal stem cells and exosomes and its application in tissue repair

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1 Preparation of Polypeptides Promoting Cell Proliferation in a High Glucose Environment

[0037] Hacat human immortalized keratinocytes (Cat. No. YCL-0090, Yipu Biology) and NHDF (Normal Human Skin Fibroblasts) (Cat. No.: HTX2167, Shenzhen Haodi Huatuo Biotechnology Co., Ltd.) were used as target cells. The peptide library constructed by the inventors was used to conduct specific screening of polypeptides that promote cell proliferation at ℃, and a total of three polypeptides that can simultaneously promote the proliferation of two types of cells in a high-glucose environment were obtained, which were named GTC-2, GTC-15, and GTC-15 respectively. GTC-29, the dodecapeptide was synthesized by Beijing Zhongke Yaguang Biotechnology Co., Ltd. and labeled with FITC, and the purity of the peptide was >95%.

Embodiment 2G

[0038] Example 2 Detection of GTC-2 polypeptide activity

[0039] HaCat human immortalized keratinocyte culture medium: culture cells with RPMI1640 medium containing 10% fetal bovine serum and 50mmol / L glucose, and add 1% penicillin-streptomycin solution with a volume of 10000U / ml-10000μg / ml .

[0040] Normal human skin fibroblast culture medium: culture cells with 10% fetal bovine serum, 50mmol / L MEM medium, add 1% volume of 10000U / ml-10000μg / ml penicillin-streptomycin, 1% volume sodium pyruvate, 5mML glutamine.

[0041] Take the cryopreservation tubes containing HaCat human keratinocytes and human normal fibroblasts out of the liquid nitrogen and quickly place them in a 37°C water bath to melt them quickly. After the liquid in the tubes is completely melted, suck out the cell fluid Put 4ml of complete culture solution in a centrifuge tube, centrifuge at 1000rpm for 5 minutes, then discard the supernatant with a straw of an autoclave, mix the precipitated cells evenly with 5m...

Embodiment 3

[0043] Embodiment 3 GTC-2 polypeptide influences PCR detection on corresponding gene of cell

[0044] The two kinds of cells treated with HaCat human immortalized keratinocyte control group and low concentration group in Example 2 were respectively extracted RNA for reverse transcription polymerase chain reaction, reverse transcription system 20 μl, containing template RNA; according to TaKaRaRNALAPCRTMKit reagent The box provides instructions for operation. The reaction conditions are: 55°C for 30 minutes, 99°C for 5 minutes, and 5°C for 5 minutes; the PCR reaction system is 15 μl, containing 3 μl reverse transcription reaction product, 0.9 μl MgCl 2 (25mM), 1.2μl PCR reaction buffer, 0.075μl Taq polymerase, 0.3μl upstream primer, 0.3μl downstream primer, with H 2 O(1‰DEPC) complements the system. The PCR reaction conditions were: pre-denaturation at 95°C for 3 minutes, denaturation at 95°C for 1 minute, annealing at 58°C for 55 seconds, extension at 72°C for 40 seconds, 35 ...

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Abstract

The invention relates to a composition comprising mesenchymal stem cells and exosomes and its application in tissue repair. Combining the repair peptide provided by the present invention with adipose stem cells and epidermal stem cell exosomes can effectively repair the ulcer model of diabetic mice and significantly promote the repair of diabetes loss, which has good application prospects and application value.

Description

technical field [0001] The present invention relates to the biological field, in particular to a composition comprising mesenchymal stem cells and exosomes and its application in tissue repair. Background technique [0002] Chronic skin ulcer caused by trauma is one of the reasons for diabetic patients to be hospitalized. Its condition is complex and easy to relapse, which can lead to serious complications. The treatment is relatively difficult, the treatment methods are limited, and the incidence rate is increasing year by year. It is a difficult problem that is eager to overcome in clinical practice . It is known that wound healing is a complex and delicate process involving a variety of cells and molecules. Current studies have shown that more than hundreds of physiological factors are associated with poor wound healing in diabetes, such as decreased growth factors, decreased angiogenesis, low number and impaired function of macrophages, abnormal collagen deposition, and...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K38/16A61K8/64A61K8/99A61P3/10A61P17/02A61P39/00A61Q19/00C07K14/00C12N5/0775A61K35/28
CPCA61K8/64A61K8/99A61K35/28A61K38/16A61Q19/00A61P3/10A61P17/02A61P39/00C07K14/00C12N5/0667C12N5/0668
Inventor 韩卫星曹燕黄玉燕张涛陈博
Owner 广东肽诺干细胞生物科技有限公司
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