Application of endometrium stem cell in preparation of medicine for curing liver fibrosis

A technology of endometrial stem cells and liver fibrosis, applied in animal cells, drug combinations, vertebrate cells, etc., can solve the problems of donor shortage and high cost, and achieve the effect of improving liver function and reducing the degree of liver fibrosis

Pending Publication Date: 2016-12-21
HANGZHOU S EVANS BIOSCI LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Orthotopic liver transplantation is an effective method for the treatment of end-stage liver cirrhosi...

Method used

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  • Application of endometrium stem cell in preparation of medicine for curing liver fibrosis
  • Application of endometrium stem cell in preparation of medicine for curing liver fibrosis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] Example 1: Preparation of chronic liver injury mouse model

[0016] Utilize 10% (volume concentration) carbon tetrachloride (CCl 4 , the solvent is olive oil) to induce chemical liver injury to eight-week-old ICR mice (about 25 grams in body weight), intraperitoneal injection, the dose is 1ml CCl 4 / kg body weight, 2 times a week, continuous injection for 4 weeks, to establish the liver fibrosis model, and the injected mice were fed normally in a sterile environment.

Embodiment 2

[0017] Embodiment 2: the preparation of serum preparation

[0018] Serum preparation: Blood was collected from the orbital blood method of liver fibrosis model mice as described in Example 1, or venous blood was collected from patients with liver fibrosis, kept in a water bath at 37°C for 30 minutes, centrifuged at 3500rpm for 10 minutes, and the supernatant was taken at -20 ℃ frozen for later use.

[0019] Serum preparation preparation: DMEM culture fluid + penicillin + 100U / ml streptomycin + volume concentration 1% amphotericin B + volume concentration 1% glutamine + volume concentration 20% fetal bovine serum, add volume final concentration of 1 to 5% The above-mentioned serums are ready-to-use and ready-to-use.

Embodiment 3

[0020] Example 3: Endometrial Stem Cell Preparation

[0021] (1) Isolation and culture of endometrial stem cells

[0022] Collect menstrual blood samples from donors for routine microbial and infectious disease pathogen safety testing, use 1.0779 / ml Ficoll for density gradient centrifugation, separate mononuclear cells, wash the isolated mononuclear cells 3 times with normal saline, and use DMEM culture medium (add 100U / ml penicillin+100U / ml streptomycin+volume concentration 1% amphotericin B+volume concentration 1% glutamine+volume concentration 20% fetal bovine serum) adjust cell density to (3~5)×10 6 cells / ml, seeded on the bottom area of ​​75cm 2 culture flask at 37°C, 5% CO 2 Cultivate in an incubator; after 48 hours, wash off non-adherent cells, continue to culture until about 80% confluence, digest with 0.25% trypsin-EDTA, and pass passage.

[0023] (2) Preparation of endometrial stem cell preparation

[0024] Take the endometrial stem cells that have grown to about...

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Abstract

The invention discloses an application of an endometrium stem cell in preparation of a medicine for curing liver fibrosis, wherein the endometrium stem cell is a fibroblast-like cell growing by adhering to a wall, expresses CD29, CD44, CD73 and Cd90, and does not express CD34, CD45, CD117 and HLA-DR. In the application, the endometrium stem cell is convenient to acquire, the preparation of an endometrium stem cell is simple, the infusion technique is simple and convenient, and is easy to operate, the effects of improving the liver function and reducing liver fibrosis are remarkable, and the application is of great clinical significance and has a wide application prospect.

Description

(1) Technical field [0001] The invention relates to the field of liver fibrosis treatment, in particular to the application and preparation method of endometrial stem cells in the treatment of liver fibrosis. (2) Background technology [0002] Hepatic fibrosis is the main pathological feature of chronic liver diseases caused by hepatitis, chronic alcoholism, metabolism and heredity. Its later stage leads to liver cirrhosis and liver cancer, which seriously threatens health and life. my country is a big country of hepatitis. Among patients with hepatitis B, especially chronic active hepatitis B, 10%-15% of patients may develop post-hepatitis cirrhosis within 5-10 years. Orthotopic liver transplantation is an effective method for the treatment of end-stage liver cirrhosis, but factors such as lack of donors, high cost, and immune rejection limit its widespread implementation. Therefore, it is still very urgent to develop effective drugs and methods for the treatment of liver ...

Claims

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Application Information

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IPC IPC(8): A61K35/33A61P1/16C12N5/0789
CPCA61K35/33C12N5/0634
Inventor 戴玲华张强
Owner HANGZHOU S EVANS BIOSCI LTD
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