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Composition and application thereof

A composition and drug technology, applied in the field of cells, can solve problems such as the unclear understanding of heterogeneous cell proliferation, differentiation, and changes in lifespan immune regulation functions, so as to shorten tissue adaptability, avoid differentiation uncertainty, and prepare Easy to prepare

Inactive Publication Date: 2017-02-22
GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, the understanding of the proliferation, differentiation, lifespan of allogeneic cells in the body after infusion and the changes in immune regulation function brought about by this is not very clear, and the problems arising from clinical application have fallen into a bottleneck period

Method used

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  • Composition and application thereof
  • Composition and application thereof
  • Composition and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0076] Primary culture of umbilical cord stem cells (or purchased from the market):

[0077] Take the umbilical cord of a healthy full-term caesarean section baby, clean it aseptically in an ultra-clean bench, peel off the umbilical cord amniotic membrane and arteries and veins, take out Wharton’s glue, cut it into pieces with surgical scissors, resuspend it with DMEM+10% FBS complete medium and inoculate to a 15cm dish, transfer to 5% CO 2 , cultivated in an environment of 37 degrees Celsius.

[0078] Replenish after 5 days until the cells climb out, discard the tissue pieces, supplement with new complete medium and continue to culture, when the cells form larger clones, they can be passaged

[0079] Subculture of umbilical cord stem cells:

[0080] ① Take subcultured P0 cells, discard the original culture medium in the dish, add 10mL PBS buffer solution to gently wash the cell growth surface, and discard the washing solution.

[0081] ②Add 2mL of 0.25% trypsin, shake the ...

Embodiment 2

[0092] The preparation of embodiment 2 compositions:

[0093] ① Take out exosomes from -20°C, dissolve them in an environment of 4°C, and mix according to the composition: exosomes + normal saline. Mix well and store at 4°C for later use. The content of exosomes is 5×10 7 The total amount of exosomes extracted from cells.

[0094] ②Resuspend the hepatic cells with the preparation of exosomes, mix well and store in an environment at 4°C for later use. The content of hepatoid cells was 1×10 6 cells.

[0095] Preparation specific steps:

[0096] ①Add 100 μL 5×10 7 The total amount of exosomes extracted from cells.

[0097] ②Resuspend 1×10 exosome suspension 6 cells of hepatic-like cells.

[0098] ③ Add physiological saline to a final volume of 5 mL.

Embodiment 3

[0100] ① Take out exosomes from -20°C, dissolve them in an environment of 4°C, and mix according to the composition: exosomes + normal saline. Mix well and store at 4°C for later use. The content of exosomes is 1×10 7 The total amount of exosomes extracted from cells.

[0101] ②Resuspend the hepatic cells with the preparation of exosomes, mix well and store in an environment at 4°C for later use. The content of hepatoid cells was 5×10 6 cells.

[0102] Preparation specific steps:

[0103] ① Add volume of 100μL 1×10 7 The total amount of exosomes extracted from cells.

[0104] ②Resuspended with exosome suspension 5×10 6 cells of hepatic-like cells.

[0105] ③ Add physiological saline to a final volume of 5 mL.

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Abstract

The invention relates to the technical field of cells, in particular to a composition and an application thereof. The composition comprises an umbilical cord stem cell exosome and a hepatocyte-like cell. The composition adopts the unit of the umbilical cord stem cell exosome and the hepatocyte-like cell to conduct tissue local injection, thus loss of exosome caused by hemokinesis can be avoided. The exosome can repair a damaged tissue directly, strengthen a resistant ability of an undamaged tissue, and shorten sharply repairing time of the tissue. The exosome preparation does not have immunogenicity, thus can be applicable to any individual. The hepatocyte-like cell can shorten adaptation of the tissue, perform functions directly under a microenvironment, and shorten cure period. The application of the exosome is safer compared with an direct application of the hepatocyte-like cell, and a method of non-cell therapy can be used. The exosome can be stored for a long time after being extracted, and is more convenient in preparation preparing during the application. Operations such as culturing cells again in a cell therapeutic scheme are avoided. By using the hepatocyte-like cell, the disadvantage of uncertainty of stem cell differentiation can be avoided.

Description

technical field [0001] The invention relates to the field of cell technology, in particular to a composition and its application. Background technique [0002] Liver disease is a common clinical disease and frequently-occurring disease, which seriously endangers human health. A large number of liver disease patients died due to the lack of ideal and effective treatment. At present, methods such as liver transplantation are used to delay the disease; the type of liver is low, and the source is few; the recurrence rate is high, and the long-term survival rate of patients is low. [0003] Bone marrow mesenchymal stem cells are a kind of pluripotent stem cells with the potential of self-proliferation and differentiation. They are widely used in liver diseases because of their plasticity, convenient material acquisition, low immunogenicity, and easy transfection and expression of foreign genes. Research. Recent studies have shown that in a suitable microenvironment, bone marro...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/51A61P1/16A61K35/407
CPCA61K35/407A61K35/51A61K2300/00
Inventor 葛啸虎陈海佳王一飞卢瑞珊王小燕
Owner GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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