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

Active Publication Date: 2012-07-11
中源协和生物细胞存储服务(天津)有限公司
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Problems solved by technology

Therefore, the application of induced hepatocytes is limited and cannot meet a large number of needs, such as replacement therapy for liver diseases and screening of cell drugs
The current technology of differentiation of mesenchymal stem cells (MSCs) into hepatocytes has certain defects and deficiencies, such as high differentiation cost, low differentiation efficiency, etc., which is not conducive to the industrialization of cell differentiation process, future drug screening and clinical use

Method used

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  • Inducing method for directional differentiation of human umbilical cord mesenchymal stem cells into liver cells
  • Inducing method for directional differentiation of human umbilical cord mesenchymal stem cells into liver cells
  • Inducing method for directional differentiation of human umbilical cord mesenchymal stem cells into liver cells

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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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Abstract

The invention provides an induced culture method of liver cells, particularly provides a method for directional induced differentiation of human umbilical cord mesenchymal stem cells into liver cells and more particularly relates to a method for staged directional induced differentiation of human umbilical cord mesenchymal stem cells into liver cells by extracting human umbilical cord mesenchymalstem cells from abandoned umbilical cords through a cell climbing sheet technique, culturing the extracted stem cells, and adding an epidermal growth factor, dexamethasone, a liver cell growth factor, oncostatin and the like.

Description

technical field [0001] The induction method can utilize human umbilical cord mesenchymal stem cells to induce directed differentiation into liver cells. Background technique [0002] At present, embryonic stem cells and bone marrow mesenchymal stem cells are used to induce differentiation into hepatocytes. However, embryonic stem cells are difficult to culture and expand, have strong tumorigenicity, and have ethical issues; bone marrow mesenchymal stem cells have limited sources, are difficult to expand, and have certain damage to the donor. Therefore, the application of induced hepatocytes is limited and cannot meet a large number of needs, such as the replacement therapy for liver diseases and the screening of cell drugs. The current technology of differentiation of mesenchymal stem cells (MSCs) into liver cells has certain defects and deficiencies, such as high differentiation cost, low differentiation efficiency, etc., which is not conducive to the industrialization of ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/071
Inventor 王汉裕刘拥军刘广洋朱德琳赵婷婷
Owner 中源协和生物细胞存储服务(天津)有限公司
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