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Application of exosome derived from human mesenchymal stem cells to resistance to tissue fibrosis and scar forming

A technology of stem cells and exosomes, applied in drug research and development, can solve the problems of limited clinical application, difficult storage, safety problems, etc., and achieve the effect of convenient resource acquisition, easy absorption, and inhibition of tissue and organ fibrosis.

Inactive Publication Date: 2016-04-13
SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, MSC itself is not easy to store and its activity declines sharply over time, and there are safety problems after transplantation into the body. Long-term use may even have potential tumorigenic potential, which greatly limits clinical applications. Looking for alternatives to MSC in tissue damage repair Biomaterials or stem cell products that antagonize fibrosis and scar hyperplasia are urgently needed

Method used

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  • Application of exosome derived from human mesenchymal stem cells to resistance to tissue fibrosis and scar forming
  • Application of exosome derived from human mesenchymal stem cells to resistance to tissue fibrosis and scar forming
  • Application of exosome derived from human mesenchymal stem cells to resistance to tissue fibrosis and scar forming

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1: Preparation process of exosomes derived from mesenchymal stem cells

[0030] (1) Isolation, culture and expansion of mesenchymal stem cells:

[0031] 1) Take the fresh umbilical cord of a newborn, wash it repeatedly with phosphate buffer, soak it in L-DMEM containing 1% penicillin and streptomycin for about 5 minutes, and cut it into tissue pieces with a diameter of about 1.5mm; Bovine serum L-DMEM nutrient solution, 5% CO2, 37°C saturated humidity culture; after 7-10 days, mesenchymal stem cells can be seen climbing out of the tissue, after removing the tissue pieces, digest with 0.25% trypsin for subculture. Identification results such as figure 1 shown. It can be seen that umbilical cord mesenchymal stem cells highly express CD29, the positive rate is 99.7%; CD44, the positive rate is 94.4%, do not express CD14, the positive rate is 0.2%; do not express CD45, the positive rate is 3.1%.

[0032] 2) Take the bone marrow of a healthy person, expose the bon...

Embodiment 2

[0045] Example 2: Cytological study on the ability of exosomes secreted by umbilical cord mesenchymal stem cells to regulate the differentiation of fibroblasts into myofibroblasts

[0046] (1) Isolation, culture and expansion of fibroblasts: Take normal human skin tissue, wash it repeatedly with phosphate buffer saline, soak it in H-DMEM containing 1% penicillin and streptomycin for about 5 minutes, cut it into a diameter of about Tissue blocks of 1.5mm size were pasted in a culture dish; cultured in H-DMEM nutrient solution containing 10% fetal bovine serum, 5% CO2, and 37°C saturated humidity; after 7-10 days, fibroblasts and epidermal cells could be seen crawling out of the tissue, After removing the tissue pieces, digest with 0.25% trypsin for about 1 minute according to the digestion time required for fibroblasts, and subculture after removing epidermal cells.

[0047] (2) Establish a cell model of fibroblast differentiation into myofibroblasts: spread fibroblasts on a 6-we...

Embodiment 3

[0075] Example 3. Exosomes derived from mesenchymal stem cells alleviate liver fibrosis in rats

[0076] (1) Rat liver fibrosis model is established: CCl is made into 50% olive oil agent, and each rat in the treatment group is injected intraperitoneally by 1ml / kg body weight, three times in the first, twice a week after the second week, and simultaneously Feed 10% alcoholic beverages for six weeks.

[0077] (2) Exosome intervention therapy derived from mesenchymal stem cells:

[0078] Rats with hepatic fibrosis were randomly divided into 2 groups, 10 in each group.

[0079] Treatment method: exosome group: injecting exosomes with CCl 4 After the second week, start to treat once every three days, inject the present invention into the tail vein, and the dose is 150ug / kg for 3 weeks. Blank control group: the treatment time was the same as that of the exosome group, once every three days, the same volume of normal saline was injected into the tail vein for a total of 3 weeks. ...

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Abstract

The invention belongs to the technical field of molecular biology and relates to application of an exosome derived from human mesenchymal stem cells to preparation of medicines for antagonizing scars or tissue and organ fibrosis. According to the application, the high-purity and high-bioactivity exosome is obtained by subjecting healthy human umbilical cords to primary in-vitro culture, collecting liquid supernatant after amplification, performing low-temperature ultracentrifugation, purifying and the like; the exosome is used individually or added into medicines, health care products and cosmetics according to a proportion to be developed into a product with a bioactivity function, thereby being capable of regulating organism cell signal transduction effectively, inhibiting fibrocyte differentiation and then antagonizing the tissue and organ fibrosis and scar forming. Through experimental verification, the exosome can play a remarkable role in repairing promotion in hypertrophic scar after skin injury, particularly pathological fibrosis proliferation such as scar contracture of the joint parts, hepatic fibrosis and cardiac fibrosis, and is high in safety, little in toxicity and convenient to storage and transport, thereby being wide in application prospect.

Description

technical field [0001] The invention belongs to the fields of drug research and development, cell biology and molecular biology. It involves exosomes derived from human umbilical cord mesenchymal stem cells, which can be used in a variety of situations, including antagonizing tissue and organ fibrosis, excessive scar formation, and scar adhesion of skin and joints. Background technique [0002] Myofibroblasts (myofibroblasts) are highly differentiated cells from fibroblasts expressing smooth muscle actin (α-SMA), which participate in both fiber formation and extracellular matrix remodeling in the fibrosis cascade. stages (VanDeWaterL, VarneyS, TomasekJJ, Mechanoregulation of the MyofibroblastinWoundContraction, Scarring, and Fibrosis: Opportunities for New Therapeutic Intervention. 2013May;2(4):122-141). Tomasek (TomasekJJ, GabbianiG, HimB, eta1.Myofibmblasandmechalno-regulationofconnectivetissueremodelling.NatRevMolCellBiol.2002May; 3(5):349-63.) etc. pointed out that myof...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/28A61K9/00A61K9/06C12N5/0775A61P17/02A61P1/16A61P21/00A61P9/00A61P43/00
CPCA61K9/0019A61K9/06A61K35/28C12N5/0665
Inventor 方硕刘厚奇王越邢新薛春雨杨超章云童毕宏达戴海英严冰浩
Owner SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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