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Method for stem cell differentiating mesenchymal stem cells into bile duct cells

A technology of stem cells and bile duct cells

Inactive Publication Date: 2019-01-11
SECOND AFFILIATED HOSPITAL SECOND MILITARY MEDICAL UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Many studies have reported that mesenchymal stem cells can be induced to differentiate into hepatocytes, but there is no report on the differentiation of mesenchymal stem cells into cholangiocytes

Method used

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  • Method for stem cell differentiating mesenchymal stem cells into bile duct cells
  • Method for stem cell differentiating mesenchymal stem cells into bile duct cells
  • Method for stem cell differentiating mesenchymal stem cells into bile duct cells

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Embodiment 1

[0024] 1. Reagents

[0025] 1.1 Cell culture medium

[0026] 1. DMEM medium (Invitrogen / GIBCO, #11995065)

[0027] 2. Fetal bovine serum (FBS)

[0028] 3. IMDM medium (Invitrogen / GIBCO, #12440053)

[0029] 4. William’s E culture medium (without glutamine) (Invitrogen / GIBCO, #12551-032)

[0030] 5.1× Phosphate Buffer Saline (PBS) (without Ca 2+ or Mg 2+ ) (Invitrogen / GIBCO, #10010031)

[0031] 6. 0.25% trypsin-EDTA digestion solution (Invitrogen / GIBCO, #25200056)

[0032] 7. Matrigel (Matrigel, Corning, #356234)

[0033] Type 8.1 Human Collagen (Corning, #354265)

[0034] 1.2 Cell differentiation medium

[0035] 1. Human bone marrow mesenchymal stem cell (hMSC) culture medium:

[0036] 50ml DMEM culture solution containing 10% FBS, 10ng / mL human fibroblast growth factor (Peprotech, #100-18B-100ug), 5ug / mL heparin.

[0037] 2. Serum-free medium:

[0038] 49.985ml IMDM culture solution containing 20ng / ml human epidermal growth factor (Gibco, #PHG0311) (dissolve 100ug ...

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Abstract

The invention relates to a method for differentiating mesenchymal stem cells into bile duct cells. The method comprises the following steps: (1) differentiating human mesenchymal stem cells into endodermal stem cells (cell differentiation culture; and (2) carrying out maturation differentiation on the endodermal stem cells, so as to obtain the bile duct cell (cell maturation culture). According toa ''two-step differentiation method'' for inducing the differentiation of the mesenchymal stem cells under a specific culture condition, the human mesenchymal stem cells is successfully differentiated into the functional bile duct cells, the stem cells and other cells do not need to be subjected to co-culture or genetic recombination, and the induced differentiation is carried out only based on the specific culture condition. The bile duct cells are easily available, have no immunoreaction and tumorigenesis property, can be rapidly proliferated in vitro and are hopeful of being applied to clinical drug screening, manual biliary tract reconstruction and the like.

Description

technical field [0001] The invention belongs to the field of differentiated cells, in particular to a method for differentiating mesenchymal stem cells into bile duct cells. Background technique [0002] Ideal stem cells that can be used for clinical treatment or preclinical research generally have the following characteristics: easy to obtain, no immune response, rapid proliferation in vitro, no tumorigenesis, and long survival time after transplantation. In recent years, studies have reported that embryonic stem cells and induced pluripotent stem cells can be differentiated into cholangiocytes in vitro, but the disadvantage lies in the uncontrollable proliferation and tumorigenicity of these cells after transplantation. In addition, there is a certain degree of specificity in the genetics and epigenetics of induced pluripotent stem cells, so such cells are not suitable for biological and pharmacological studies involving reprogramming and transcription mechanisms. No othe...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071C12N5/0775
CPCC12N5/0602C12N2500/30C12N2500/33C12N2500/38C12N2501/11C12N2501/115C12N2501/12C12N2501/2306C12N2501/33C12N2501/39C12N2501/91C12N2506/1353
Inventor 王昊陆李新星梁晓雯胡志前高文超
Owner SECOND AFFILIATED HOSPITAL SECOND MILITARY MEDICAL UNIV
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