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Hepatocyte-like cells from humanized adipose-derived stem cells and preparation method and application thereof

A technology of hepatocyte-like cells and adipose stem cells, applied in the field of stem cells and biomedicine, can solve the problems of inconsistent and unknown liver differentiation ability

Pending Publication Date: 2019-05-14
博品(上海)生物医药科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the liver differentiation ability of stem cells from different tissue sources is not consistent. It is still unknown which tissue-derived stem cells are more likely to differentiate into liver cells and are more conducive to the treatment of liver failure after differentiation.
In addition, how to hepatically differentiate specific tissue-derived stem cells into highly mature hepatocyte-like cells that can be used for the treatment of acute liver failure is also unknown

Method used

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  • Hepatocyte-like cells from humanized adipose-derived stem cells and preparation method and application thereof
  • Hepatocyte-like cells from humanized adipose-derived stem cells and preparation method and application thereof
  • Hepatocyte-like cells from humanized adipose-derived stem cells and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0123] Example 1. Morphology and identification of hASCs and hUCMSCs

[0124] Morphological identification of hASCs and hUCMSCs was performed as described in "Materials and Methods".

[0125] The results were as follows: three days after the primary inoculation of hASC and hUCMSC, cell colonies were formed, and the cells in the colonies were long spindle-shaped, uniform in size, arranged closely and growing radially; the morphology and proliferation speed of P5, P10, and P15 generation cells had no obvious changes ( See figure 1 A). Flow cytometry detection of P3 generation hASC and hUCMSC: CD90, CD44, CD73, and CD105 were all positively expressed, while CD34, CD45, CD11b, CD19, and HLA-DR were negatively expressed (see figure 1 B). After two weeks of adipogenic induction of hASCs and hUCMSCs, oil red O staining showed that there were a large number of translucent red lipid droplets with good refraction in hASCs cytoplasm, while only a small amount of fine red stained parti...

Embodiment 2

[0126] Example 2. Differentiation of hASC and hUCMSC to hepatocytes

[0127] Hepatic differentiation of hASCs and hUCMSCs was performed as described in "Materials and Methods".

[0128] The results were: the results of fluorescent quantitative PCR showed that compared with the hUCMSC induction group, the expression of ALB, AFP, HNF1α, HNF4α, and CYP3A4 mRNA were significantly enhanced in the hASC induction group, and the expression levels increased with the induction time (see figure 2 ).

Embodiment 3

[0129] Example 3. Alb and AFP cell immunofluorescence observation

[0130] The expression levels of Alb and AFP in hASC-induced cells and hUCMSC-induced cells were detected according to "Materials and Methods".

[0131] The results were: in the hASCs induction group, the expression levels of Alb and AFP increased with the prolongation of the induction time, and the expression level of Alb increased significantly on the 11th day (such as image 3 shown). However, in the hUCMSC induction group, the expression of ALB and AFP was not significantly increased (such as Figure 4 shown).

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Abstract

The invention provides a preparation method of hepatocyte-like cells prepared from humanized adipose-derived stem cells and an obtained hepatocyte-like cell population. The method includes the steps that in a specific induced differentiation step, a specific cell factor combination is used, and in this way, the highly-mature hepatocyte-like cells are obtained from the humanized adipose-derived stem cells and then used for treating an acute hepatic failure.

Description

technical field [0001] The invention relates to the fields of stem cells and biomedicine. Specifically, the present invention relates to human-derived adipose stem cell (Human-Adipose Derived Stem Cell, hADSC)-derived hepatocyte like cell (HLC) and its preparation method and application in the treatment of acute liver failure and other diseases. Background technique [0002] Liver failure is a serious threat to human health. Orthotopic liver transplantation is currently the best treatment, but it is expensive and limited by the source of liver. Stem cells have strong self-renewal ability and multi-directional differentiation potential across germ layers, and have become a hot spot in the treatment of liver diseases such as acute liver failure. [0003] It has been confirmed that human mesenchymal stem cells (Mesenchymal stem cells, hMSCs) can be induced to differentiate into hepatocyte-like cells (Hepatocyte like cells, HLCs) by adding a combination of several cytokines to ...

Claims

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Application Information

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IPC IPC(8): C12N5/071A61P1/16
Inventor 徐俊傅青春金银鹏高正良李洪超王晢
Owner 博品(上海)生物医药科技有限公司
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