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Method for efficiently inducing human umbilical cord mesenchymal stem cells to be differentiated into hepatocytes and application

A technology of mesenchymal stem cells and human umbilical cord, applied in the field of stem cells and regenerative medicine, can solve the problems of immature liver cell function, low yield of target cells, shorten the induction period, etc., and achieve mature cell function, high induction efficiency, and abundant sources Effect

Active Publication Date: 2021-12-14
NO 1 THE PEOPLES HOSPITAL HUAIAN CITY
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Problems solved by technology

However, at this stage, the function of the differentiated hepatocytes induced by the above methods is immature, and the yield of target cells is low, and the clinical application of transplanted cells needs to shorten the induction period as much as possible. The above induction methods usually take at least one month. The method is difficult to meet the clinical needs, so it is of great significance to develop a new method that can efficiently induce HuMSCs into mature hepatocytes in a short time

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  • Method for efficiently inducing human umbilical cord mesenchymal stem cells to be differentiated into hepatocytes and application
  • Method for efficiently inducing human umbilical cord mesenchymal stem cells to be differentiated into hepatocytes and application
  • Method for efficiently inducing human umbilical cord mesenchymal stem cells to be differentiated into hepatocytes and application

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Embodiment Construction

[0036] Combine below Figure 1-Figure 11 The relevant steps and technical advantages of the present invention are further described in detail.

[0037] 1. Acquisition of human umbilical cord mesenchymal stem cells

[0038] After the fresh umbilical cord is obtained, put it into sterile normal saline, separate the umbilical cord Huatong jelly into the ultra-clean bench, put it into the bottom of the culture bottle for adherent culture, add L-DMEM medium containing 10% fetal bovine serum, and place in 5% CO 2 , In a 37 ℃ incubator, observe the growth of spindle cells around the tissue block after 4-5 days, remove the tissue block and continue to add medium for culture, and pass passage after the cells reach 70-80% confluence.

[0039] 2. Preparation of Differentiated Cells

[0040] The 4th generation human umbilical cord mesenchymal stem cells were inoculated in culture flasks with a cell concentration of 1×10 8 / L, the culture bottle is L-DMEM medium containing 10% fetal bov...

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Abstract

The invention discloses a method for efficiently inducing differentiation of human umbilical cord mesenchymal stem cells into hepatocytes and an application. The method comprises the following steps: transfecting constructed human umbilical cord mesenchymal stem cells stably expressing long-chain non-coding RNA LINC5657, and a culture medium containing cytokines HGF, bFGF, EG and OSMby lentivirus. The hepatocyte induced differentiation culture medium is replaced once every three days, the hepatocyte-like cells (iHeps) with the hepatocyte morphology and function can be obtained only in about 21 days, the cell function is more mature than that of a traditional method, and a reliable cell source can be provided for clinical hepatocyte transplantation.

Description

technical field [0001] The invention belongs to the technical field of stem cells and regenerative medicine, and mainly relates to a method for efficiently differentiating human umbilical cord mesenchymal stem cell lines overexpressing LINC5657 into hepatocytes in a culture medium containing inducers HGF, EGF, bFGF and OSM. Background technique [0002] Various end-stage liver diseases often lead to liver failure, causing great harm to human health, and are one of the main causes of death worldwide. At present, liver cell transplantation is considered to be one of the important methods for the treatment of acute and chronic liver failure, but the lack of enough donors limits its wide application in clinical practice. [0003] With the rapid development of stem cell technology in recent years, adult stem cells provide an unlimited source of primary cells for in vitro research. Among adult stem cells, mesenchymal stem cells have the most application prospects, which have the ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071C12N15/867C12N5/10A61K35/407A61P1/16
CPCC12N5/067C12N15/86A61K35/407A61P1/16C12N2501/115C12N2501/11C12N2501/12C12N2501/237C12N2740/15043C12N2800/107C12N2501/999Y02A50/30
Inventor 禹亚彬祁付珍王丹丹
Owner NO 1 THE PEOPLES HOSPITAL HUAIAN CITY
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