Preparation method of extracellular matrix for promoting differentiation of bone marrow stem cells into liver cells

A technology of bone marrow stem cells and liver cells, applied in the field of biomedicine, can solve the problems of high cost and cumbersome steps, achieve cost reduction, facilitate popularization and application, and solve the effect of immune rejection of cell transplantation

Inactive Publication Date: 2018-09-18
广东芙金干细胞再生医学有限公司
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Problems solved by technology

[0009] In summary, because the existing hepatocyte differentiation method is cumbersome and expensive, and cannot solve the problem of how to obtain mature and usable hepatocytes without immune rejection from the perspective of clinical application, it is necessary to develop a method with simple steps and materials An economical and clinically useful method of hepatocytes for the treatment of liver diseases is necessary

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  • Preparation method of extracellular matrix for promoting differentiation of bone marrow stem cells into liver cells
  • Preparation method of extracellular matrix for promoting differentiation of bone marrow stem cells into liver cells

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Embodiment 1

[0036] Step 1: Prepare porcine liver extracellular matrix

[0037] Whole fresh livers were harvested from adult minipigs weighing 60kg to 80kg under anesthesia. The hepatic artery and hepatic bile ducts were ligated, leaving only the vena cava open as an outlet. After the catheter was inserted into the portal vein, immediately use a peristaltic pump (model such as Millipore, Billerica, MA, USA) to perfuse the pig liver with 20 L of sterile PBS solution containing 5000 IU / L heparin at a rate of 100 mL / min to remove intrahepatic blood.

[0038] Step by step at 4°C with 150L of pre-cooled 0.01% sodium dodecyl sulfate (SDS) solution, 150L of pre-cooled 0.1% SDS solution, 50L of pre-cooled 1% SDS solution and 50L of 1% pre-cooled Triton Pig livers were perfused with X-100 to decellularize them. Finally, the decellularized liver was perfused with 100 L of distilled water to remove residual surfactant and cellular debris. If quality testing is required, after the decellularization...

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Abstract

The embodiments of the invention provide a preparation method of an extracellular matrix for promoting differentiation of bone marrow stem cells into liver cells and relates to the field of biomedicine. The prepared extracellular matrix has the advantages of adjusting the micro-environment of bone marrow mesenchymal stem cells and inducing differentiation of the bone marrow mesenchymal stem cellsinto mature liver cells. The extracellular matrix can be used for studying liver disease mechanisms and screening drugs in scientific research and preparing liver cells before clinical liver cell transplantation for liver disease treatment. The preparation method of the extracellular matrix for promoting differentiation of the bone marrow stem cells into the liver cells comprises the following steps: step 1, preparing a porcine liver extracellular matrix; step 2, separating and culturing autologous bone marrow mesenchymal stem cells of patients; step 3, differentiating the bone marrow mesenchymal stem cells subjected to 3D culture in vitro into the liver cells; and step 4, performing function evaluation of the liver cells.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a preparation method of an extracellular matrix for promoting the differentiation of bone marrow stem cells into liver cells. Background technique [0002] About 800,000 to 1.2 million people die of liver disease every year in the world, accounting for about 1.3%-2% of the total deaths. As a hard-hit area with high incidence and high infection rate of liver disease in China, it is an urgent task to provide cost-effective means to treat liver disease. According to the pathological process and changes in liver function, liver diseases can be divided into liver fibrosis in the early and middle stages, liver cirrhosis in the end stage, and liver decompensation caused by the end stage. Taking hepatitis B as an example, antiviral drugs in the early, middle, and even end stages can inhibit virus replication. Or effectively relieve liver disease. However, due to the strong regeneration abili...

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Application Information

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IPC IPC(8): C12N5/071C12N5/0775
CPCC12N5/067C12N2500/20C12N2500/84C12N2506/1353
Inventor 刘文佳李丽雅明磊国毕焕京程蕊苹邓志宏
Owner 广东芙金干细胞再生医学有限公司
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