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
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[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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