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A stem cell preparation for treating liver fibrosis

A stem cell preparation and liver fibrosis technology, applied in animal cells, vertebrate cells, bone/connective tissue cells, etc., can solve the problems of decreased proliferation and differentiation potential, inconsistent culture programs, and restrictions on large-scale use, so as to improve liver function. Functional status, not limited by ethics, and the effect of promoting liver pathological recovery

Active Publication Date: 2019-01-01
HANGZHOU S EVANS BIOSCI LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among the various types of mesenchymal stem cells that have been isolated and cultured, the number and proliferation and differentiation potential of bone marrow mesenchymal stem cells decrease with the age of the donor, and they are susceptible to virus infection during the isolation and culture process, which directly affects their safety. , which restricts the large-scale use of
Although the safety of umbilical cord blood mesenchymal stem cells can be guaranteed, their number is small, the frequency is low, the culture scheme is not uniform, subculture is difficult, and it is not easy to obtain in large quantities

Method used

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  • A stem cell preparation for treating liver fibrosis
  • A stem cell preparation for treating liver fibrosis
  • A stem cell preparation for treating liver fibrosis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1: Preparation of Menstrual Blood Stem Cell Injection

[0023] Collect menstrual blood samples from donors for routine microbial and infectious disease pathogen safety testing, use 1.0779 / ml Ficoll for density gradient centrifugation, and separate mononuclear cells from qualified samples, wash the isolated mononuclear cells 3 times with PBS, adjust Cell density to (3-5)×10 6 cells / ml, seeded on the bottom area of ​​75cm 2 placed in a culture flask at 37°C, 5% CO 2 Expansion culture in the incubator, using DMEM medium, 1% double antibody medium by volume, 1% amphotericin B by volume, 1% glutamine by volume and 20% fetal bovine serum by volume After 48 hours of cell culture, digest with 0.25% trypsin-EDTA for 5 minutes, add an appropriate volume of complete medium to neutralize trypsin activity, and gently blow with a pipette to suspend adherent cells. The cells were washed with PBS, and the mesenchymal stem cells were reseeded in new culture flasks, and subcul...

Embodiment 2

[0026] Example 2: Establishment of mouse liver fibrosis model

[0027] ICR mice were intraperitoneally injected with CCl 4 (with lmlCCl 4 / kg dose injection containing 10% CCl 4 olive oil) to establish liver fibrosis model, injected twice a week for 4 consecutive weeks.

Embodiment 3

[0028] Example 3: Transplantation test of menstrual blood stem cells

[0029] Tail vein injection transplantation experiment: liver fibrosis model mice were divided into 3 groups, and l×10 6 cells / 100μl, 2×10 6 cells / 100μl and 4×10 6 1 cell / 100 μl cell dose, and another 100 μl PBS was injected as a control, both for one injection. Maintain CCl during transplantation 4 Inject until mice are anesthetized and sacrificed for sampling.

[0030] Evaluation of the living conditions of the mice: the mice in the control group were active, their fur was shiny, and their diets were normal. The mice in the model group had reduced activity, dry fur, and reduced food intake. The mice in the treatment group became active gradually, their fur dryness decreased, and their food intake increased.

[0031]Liver function index: On the 28th day after cell transplantation, the mice in the model group, the control group and the transplantation group were anesthetized with ether and blood was ta...

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Abstract

The invention provides a stem cell preparation for treating hepatic fibrosis, and a preparation method of the stem cell preparation. The stem cell preparation for treating hepatic fibrosis is prepared from human menstrual blood mesenchymal stem cells, a chemotactic factor CXCL8 and normal saline. The obtaining method of the menstrual blood mesenchymal stem cells is simple, free of ethical restriction, high in cell viability and high in proliferation capacity, has multi-directional differentiation potential and is simple and convenient to prepare. The menstrual blood mesenchymal stem cell preparation infusion technology provided by the invention is simple and convenient to operate; the hepatic fibrosis is treated by virtue of the menstrual blood mesenchymal stem cells; and the developed preparation method of the stem cell preparation for treating the hepatic fibrosis has very important clinical significance and wide application prospect.

Description

technical field [0001] The invention relates to the application of stem cells in disease treatment, in particular to the application of menstrual blood-derived mesenchymal stem cells in the treatment of liver fibrosis. Background technique [0002] Liver fibrosis is one of the pathological features of most end-stage liver diseases. If liver fibrosis is not treated effectively, it will eventually develop into liver cirrhosis and liver cancer, which seriously threaten the lives of patients. The essence of liver fibrosis is that collagen synthesis in the extracellular matrix increases and degradation decreases, resulting in excessive deposition of extracellular matrix in the liver, affecting the normal function of the liver. At present, it is believed that the mechanism of liver fibrosis is that liver cell injury and liver tissue inflammation activate Kupffer cells, liver cells, platelets or sinusoidal endothelial cells to secrete cytokines, lipid peroxides, etc. The substance...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/0775A61K35/28A61P1/16
Inventor 吴晓星刘广平项春生戴玲华
Owner HANGZHOU S EVANS BIOSCI LTD
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