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Use of exosomes to improve tissue repair function of mesenchymal stem cells

A technology for exosomes and epidermal stem cells, which is applied in the field of improving the tissue repair function of mesenchymal stem cells and exosomes, achieving the effects of low production cost and use cost, promoting repair, good application prospect and application value

Active Publication Date: 2021-04-27
中科天诚(深圳)集团有限公司
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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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  • Use of exosomes to improve tissue repair function of mesenchymal stem cells
  • Use of exosomes to improve tissue repair function of mesenchymal stem cells
  • Use of exosomes to improve tissue repair function of mesenchymal stem cells

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 Biotech) 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 inventor was used to screen specific peptides that promote cell proliferation at ℃, and three peptides 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 2

[0038] Example 2 Detection of GTC-15 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 MEM medium containing 10% fetal bovine serum and 50mmol / L, and 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 ce...

Embodiment 3

[0043] Example 3 PCR detection of the influence of GTC-15 polypeptide on the corresponding genes of cells

[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, and 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 (25 mM), 1.2 μl PCR reaction buffer, 0.075 μl Taq polymerase, 0.3 μl upstream primer, 0.3 μl downstream primer, in 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...

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Abstract

The present invention relates to the use of exosomes for improving the tissue repair function of mesenchymal stem cells. 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 field of biology, in particular to the use of exosomes for improving the tissue repair function of mesenchymal stem cells. 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 decreased ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K38/16A61K35/28A61K35/36A61P3/10A61P17/02A61K8/64A61K8/99A61Q19/00C07K14/00
CPCA61K8/64A61K8/99A61K35/28A61K35/36A61K38/16A61Q19/00A61P3/10A61P17/02C07K14/00A61K2300/00
Inventor 张涛陈博
Owner 中科天诚(深圳)集团有限公司
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