Inducing method for directional differentiation of human umbilical cord mesenchymal stem cells into liver cells
A technology of mesenchymal stem cells and directional induction, which is applied in the field of induction of directional differentiation of human umbilical cord mesenchymal stem cells to hepatocytes, which can solve the unfavorable cell differentiation process of industrial drug screening and clinical use, unable to meet a large number of needs, low differentiation efficiency, etc. problems, to achieve the effect of solving the source of liver cells, maintaining biological characteristics and activity, and fast cell growth
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Embodiment 1
[0013] Example 1 Obtaining Human Umbilical Cord Mesenchymal Stem Cells by Separation Technique
[0014] The umbilical cord of the newborn was taken, and the umbilical cord mesenchymal stem cells were isolated by the following steps: (1) 8g of NaCl, 0.2g of KCl, 1.15g of Na 2 HPO4, 0.2g of KH 2 PO4 was fixed to 1L with water to make a phosphate buffer solution, and the umbilical cord was washed repeatedly with the buffer solution to remove residual blood to obtain a clean umbilical cord; (2) crush the clean umbilical cord to obtain a large number of umbilical cord tissue pieces; (3) the obtained umbilical cord The umbilical cord tissue blocks were filtered through a coarse filter with a pore size of 1.5 mm, and the umbilical cord tissue blocks under the coarse filter were collected, and the large umbilical cord tissue blocks that did not meet the requirements were removed to obtain small umbilical cord tissue blocks with a diameter of 1.5 mm or less; (4) Filter the obtained sm...
Embodiment 2
[0020] Example 2 Directed induction of human umbilical cord mesenchymal stem cells into hepatocytes
[0021] The mesenchymal stem cells isolated from the umbilical cord by the slide method were subcultured to the third generation. Differentiation of MSCs into hepatocytes was induced using the phase induction method: first, the cells were treated with 2% fetal bovine serum (FBS), 10 -8 mol / L dexamethasone (DXM), 50ng / ml HGF (hepatocyte growth factor), 10ng / ml EGF (epidermal growth factor), 1% ITS (25g / L insulin, 25g / L transferrin, 25mg / L sodium selenite) in DMEM-F12 medium for induction culture for 2 weeks, then, use 2% FBS, 20ng / ml OSM (oncostatin), 10 -6 mol / L DXM, 1% ITS DMEM-F12 medium induction culture for 2 weeks. MSCs cultured in DMEM-F12 containing 10% FBS served as a negative control.
[0022] Changes in cell morphology: A series of changes occurred in the cell morphology of umbilical cord mesenchymal stem cells G09163 in the process of differentiation into hepatocy...
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