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Method for inducing differentiation of human adipose derived stromal cells into liver cells through micromolecules

A mesenchymal stem cell and small molecule technology, applied in the field of small chemical molecules to induce human adipose-derived mesenchymal stem cells to differentiate into liver-like cells, can solve the problem of long-term liver cells

Inactive Publication Date: 2020-07-31
QINGDAO UNIV
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Problems solved by technology

[0003] The purpose of the present invention is to provide a method for small molecules to induce human adipose-derived mesenchymal stem cells to differentiate into liver-like cells, which solves the problems in the prior art (1) the source of primary hepatocytes (2) the time lag of inducing functional hepatocytes in vitro long question

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  • Method for inducing differentiation of human adipose derived stromal cells into liver cells through micromolecules
  • Method for inducing differentiation of human adipose derived stromal cells into liver cells through micromolecules
  • Method for inducing differentiation of human adipose derived stromal cells into liver cells through micromolecules

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Embodiment Construction

[0027] The present invention will be described in detail below in combination with specific embodiments.

[0028] 1. Induction of Definitive Endoderm (DE) cells

[0029] (1) Select the fourth-generation hAD-MSCs human adipose-derived mesenchymal stem cells as seed cells, and digest them for later use;

[0030] (2) Cells in the experimental group and the control group were divided into 5×10 5 The density of cells / well was seeded in a 6-well plate, and hAD-MSCs medium was added to 2ml / well, and cultured for 24h;

[0031] (3) Discard the medium, wash once with PBS, add F12-DMEM Duchenne's modified Eagle medium containing 0.5% DMSO and 2% serum: nutrient mixture F-12 medium, 2ml / well, and induce for 24h;

[0032] (4) Discard the induction medium containing 0.5% DMSO, wash once with PBS, add CHIR99021 (3μM) (C 22 h 18 Cl 2 N 8 ) F12-DMEM medium containing 2% serum, 2ml / well, induced for 24h;

[0033] (5) Discard the F12-DMEM medium containing 2% serum containing 3 μM CHIR990...

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Abstract

The invention discloses a method for inducing differentiation of human adipose derived stromal cells into liver cells through micromolecules. The method comprises the steps of inducing limited endoderm cells; through induction by DE cells, obtaining liver progenitor cells; and performing induction to obtain liver mature cells. The method has the beneficial effects that hAD-MSCs taking and in-vitroculture condition is mature, can be from self adipose tissue of a patient, is free from ethical hindrances to prevent nonself cell immunological rejection, and is free from immunogenicity and high incell proliferation activity. Only about 15 days, iHeps having liver cell shape, gene expression and functions can be obtained, the induction process is quick and efficient, and the iHeps can become astable source of hepatocyte transplantation in a clinical treatment scheme.

Description

technical field [0001] The invention belongs to the technical field of stem cells and regenerative medicine, and relates to a method for inducing small chemical molecules to differentiate human adipose-derived mesenchymal stem cells into liver-like cells. Background technique [0002] Various pathogenic factors, including viral infections, toxins, autoimmune deficiencies, and genetic diseases, can impair liver function, leading to chronic liver disease or acute liver failure. About 10% of patients with end-stage liver disease die each year while waiting for a liver donor. Orthotopic liver transplantation (OLT) is an effective means of treating end-stage liver disease, however, the use of OLT is limited by the lack of donors, side effects of immunosuppressants, and ethical issues. Hepatic cell transplantation and artificial liver support have become effective methods to restore liver function and improve the survival rate of patients, but these two methods are also limited d...

Claims

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Application Information

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IPC IPC(8): C12N5/071C12N5/0775
CPCC12N5/067C12N2506/1384C12N2500/90
Inventor 尹侃许杨李霄霞
Owner QINGDAO UNIV
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