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

A kind of stem cell, directional induction technology, which is applied in the field of directional differentiation of human umbilical cord mesenchymal stem cells to hepatocytes, which can solve the problem of unfavorable cell differentiation process, industrialized drug screening and clinical use, unable to meet a large number of needs, and low differentiation efficiency. To solve the problem of liver cell source, maintain biological characteristics and activity, and achieve the effect of fast cell growth

Active Publication Date: 2011-11-23
中源协和生物细胞存储服务(天津)有限公司
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  • Application Information

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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

Examples

Experimental program
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Embodiment 1

[0013] Example 1 Obtaining Human Umbilical Cord Mesenchymal Stem Cells Using Slice Separation Technique

[0014] Take the umbilical cord of the newborn, and obtain the umbilical cord mesenchymal stem cells through the following steps: (1) mix 8g of NaCl, 0.2g of KCl, 1.15g of NaCl 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, and a clean umbilical cord was obtained; (2) the clean umbilical cord was crushed to obtain a large number of umbilical cord tissue pieces; (3) the obtained The umbilical cord tissue block is filtered through a coarse filter with a pore size of 1.5mm, the umbilical cord tissue block under the coarse filter is collected, and the large umbilical cord tissue block that does not meet the requirements is removed to obtain a small umbilical cord tissue block with a diameter less than or equal to 1.5mm; (4) Filter the obtain...

Embodiment 2

[0020] Example 2 Directed induction of human umbilical cord mesenchymal stem cells to 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 by stage induction: firstly, 2% Fetal bovin 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) DMEM-F12 medium induction culture for 2 weeks, then, use OSM (oncostatin) containing 2% FBS, 20ng / ml, 10 -6 mol / L DXM, 1% ITS DMEM-F12 culture medium induced culture for 2 weeks. MSCs cultured in DMEM-F12 containing 10% FBS served as 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 hepatocytes. The fibroblast-like morpholog...

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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 mesenchymal stem 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 hepatocytes 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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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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