Method for induced differentiation of adipose tissue-derived stromal cells into liver-like cells

Abstract

The invention provides a method for induced differentiation of adipose tissue-derived stromal cells into liver-like cells. The method comprises the following steps: washing and cleaning adipose tissue, and adding isovolumetric tissue digestive enzyme for digestion; after completion of digestion, adding a trypsin solution for digestion; after completion of digestion, carrying out centrifuging so asto remove upper-layer grease and a digestive fluid, wherein precipitates are cells of the stromal vascular fraction of the adipose tissue; resuspending the cells of the stromal vascular fraction withphysiological saline, carrying out sieving so as to remove large tissues, carrying out washing, carrying out centrifuging so as to remove a supernatant, and after washing is completed, subjecting stromal vascular cells to inoculated culture so as to obtain the adipose tissue-derived stromal cells; selecting P3-generation adipose tissue-derived stromal cells for digestion so as to obtain a single-cell suspension, and carrying out inoculated culture with a culture medium overnight; removing a serum-free proliferation culture medium, and adding a first induction culture solution for induced culture; and replacing the first induction culture solution with a second induction culture solution for induction so as to obtain differentiated liver-like cells. The method provided by the invention canobtain functional liver cells by induction.

Description

technical field [0001] The invention belongs to the field of stem cells and regenerative medicine, and in particular relates to a method for inducing the differentiation of adipose-derived mesenchymal stem cells into liver-like cells. Background technique [0002] The liver is the largest organ in the human viscera. It mainly performs metabolic functions, and plays a role in the body in deoxidizing, storing glycogen, synthesizing secretory proteins, and secreting bile to assist in fat digestion and so on. It plays an important role in human health, and liver failure can even be life-threatening. [0003] With the development of society, liver damage, including acute liver failure, various chronic liver diseases, and even liver cancer threaten the health of more and more people. Liver orthotopic transplantation is the best way to treat end-stage liver disease, but liver transplantation faces many problems, such as limited donors and severe immune rejection. The development ...

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Problems solved by technology

However, it is difficult to obtain autologous liver tissue for induction with tissue homogenate, and the homogenate of allogeneic liver tissue may even bring about cross-contamination, which limits the later clinical application; and the transfected transcription factors may cause changes in the cell genome and make cells cancerous. Potential safety hazards are also limited in clinical applications; mesenchymal stem cells can also be induced into liver-like cells by adding specific cytokines, but the ratio of cytokines is different, and the amount of various cytokines used will directly affect the induction efficiency , and at this stage, the function of liver-like cells induced by specific cytokines is limited, the yield of target cells is low, and there are still serious deficiencies

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