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Medium for inducing differentiation of adipose-derived mesenchymal stem cells into hepatocytes in vitro

A stem cell, adipose-derived technology, applied in the field of stem cell differentiation, can solve the problems of high cost, low transfection efficiency, tumorigenicity, etc., and achieve the effects of low cost, high induction efficiency and simple operation

Active Publication Date: 2019-07-16
SHENZHEN WINGOR BIO TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The current techniques for inducing MSCs to differentiate into hepatocytes include: changing the growth environment of stem cells, such as the combined use of cytokines, small molecular compounds, cell substrates, etc., but how to properly match various factors to promote the efficiency of induction of differentiation is still a problem; using gene transfer technology Transformation of cells, but there are safety issues such as tumorigenicity; protein transduction technology uses viral vectors to mediate protein gene transfer into cells, but there is a problem of low transfection efficiency
Commonly used cytokine combination culture systems include fibroblast growth factor (fibroblastg rowth factor, FGF), hepatocyte growth factor (hepatocyte growth factor, HGF) and epidermal growth factor (Epidermal Growth Factor, EGF). higher

Method used

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  • Medium for inducing differentiation of adipose-derived mesenchymal stem cells into hepatocytes in vitro
  • Medium for inducing differentiation of adipose-derived mesenchymal stem cells into hepatocytes in vitro
  • Medium for inducing differentiation of adipose-derived mesenchymal stem cells into hepatocytes in vitro

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0086] The media used to induce the differentiation of adipose-derived mesenchymal stem cells into hepatocytes in vitro are:

[0087] Digestive enzyme solution: equal volume of 0.15% type I collagenase, 0.15% hyaluronidase, 2U / ml chondroitin sulfate;

[0088] Adipose-derived mesenchymal stem cell culture medium: DMEM medium containing 10% volume fraction of fetal bovine serum;

[0089] Synchronization medium: serum-free IMDM medium;

[0090] Differentiation induction medium: IMDM medium with 15ng / ml basic fibroblast growth factor, 3% volume fraction of fetal bovine serum, 35ng / ml hepatocyte growth factor and 2.5μM retinoic acid;

[0091] Maturation medium: IMDM medium with 3% volume fraction of fetal bovine serum, 15ng / ml Oncostatin M, 2μM dexamethasone and 2% volume fraction of insulin-transferrin-selenium.

[0092] Culture of adipose-derived mesenchymal stem cells

[0093] Take adipose tissue under aseptic conditions, wash 3 times with 3 times the volume of phosphate buff...

Embodiment 2

[0102] The media used to induce the differentiation of adipose-derived mesenchymal stem cells into hepatocytes in vitro are:

[0103] Digestive enzyme solution: equal volume of 0.2% type I collagenase, 0.2% hyaluronidase, 3U / ml chondroitin sulfate;

[0104] Adipose-derived mesenchymal stem cell culture medium: DMEM medium containing 15% volume fraction of fetal bovine serum;

[0105] Synchronization medium: serum-free IMDM medium;

[0106] Differentiation induction medium: IMDM medium with 14ng / ml basic fibroblast growth factor, 2% volume fraction of fetal bovine serum, 30ng / ml hepatocyte growth factor and 2.5μM retinoic acid;

[0107] Maturation medium: IMDM medium with 3% volume fraction of fetal bovine serum, 28ng / ml Oncostatin M, 1.5 μM dexamethasone and 3% volume fraction of insulin-transferrin-selenium.

[0108] Culture of adipose-derived mesenchymal stem cells

[0109] Adipose tissue was taken under aseptic conditions, washed 4 times with 2.5 times the volume of phos...

Embodiment 3

[0117] The media used to induce the differentiation of adipose-derived mesenchymal stem cells into hepatocytes in vitro are:

[0118] Digestive enzyme solution: equal volume of 0.1% type I collagenase, 0.1% hyaluronidase, 1U / ml chondroitin sulfate;

[0119] Adipose-derived mesenchymal stem cell culture medium: DMEM medium containing 20% ​​volume fraction of fetal bovine serum;

[0120] Synchronization medium: serum-free IMDM medium;

[0121] Differentiation induction medium: IMDM medium with 12ng / ml basic fibroblast growth factor, 4% volume fraction fetal bovine serum, 25ng / ml hepatocyte growth factor and 1 μM retinoic acid;

[0122] Maturation medium: IMDM medium with 2% volume fraction of fetal bovine serum, 15ng / ml Oncostatin M, 1 μM dexamethasone and 1% volume fraction of insulin-transferrin-selenium.

[0123] Culture of adipose-derived mesenchymal stem cells

[0124] Adipose tissue was taken under aseptic conditions, washed 3 times with 2 times the volume of phosphate ...

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Abstract

The invention relates to a medium for inducing in vitro adipose-derived mesenchymal stem cells to differentiate into hepatic cells and belongs to the technical field of stem cell differentiation. In the invention, the medium comprises a digestive enzyme solution, an adipose-derived mesenchymal stem cell medium, a synchronized medium, a differentiation inducing medium and a maturation medium. The invention also provides a kit for inducing in vitro adipose-derived mesenchymal stem cells to differentiate into hepatic cells, and application of the medium and kit in inducing in vitro adipose-derived mesenchymal stem cells to differentiate into hepatic cells. The medium provided herein enables large-scale production for adipose-derived mesenchymal stem cells, induces in vitro adipose-derived mesenchymal stem cells to efficiently differentiate into hepatic cells, and is simple to operate and low in cost.

Description

technical field [0001] The invention relates to the technical field of stem cell differentiation, in particular to a culture medium for inducing adipose-derived mesenchymal stem cells to differentiate into liver cells in vitro. Background technique [0002] With the progress of tissue engineering research in recent years, stem cell transplantation has become the most promising development direction for the treatment of advanced liver disease. Mesenchymal stem cells (MSCs) are an important member of the stem cell family, and have attracted increasing attention due to their characteristics of multi-lineage differentiation potential, hematopoietic support, immune regulation and self-replication. Mesenchymal stem cells have multi-directional differentiation potential, can differentiate into various tissue cells, and then play the role of replacing and repairing tissues. The use of MSCs to induce differentiation into hepatocytes and transplantation to repair the liver has become...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/071C12N5/0775
CPCC12N5/0018C12N5/0037C12N5/067C12N2500/25C12N2500/38C12N2500/84C12N2501/115C12N2501/12C12N2501/30C12N2506/1384
Inventor 姜舒纪惜銮张芸杨顺罗朝霞
Owner SHENZHEN WINGOR BIO TECH