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Functional liver parenchyma cells and preparation method therefor

A liver parenchymal, functional technology applied in the field of stem cell differentiation

Pending Publication Date: 2021-07-23
PEKING UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no research on the use of the above-mentioned EPS cells for directed differentiation

Method used

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  • Functional liver parenchyma cells and preparation method therefor
  • Functional liver parenchyma cells and preparation method therefor
  • Functional liver parenchyma cells and preparation method therefor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0082] Example 1 Preparation of functional hepatic parenchymal cells

[0083] We first tried a protocol for direct hepatocyte differentiation using EPS cells without a pretreatment step 6 (i.e. only S2-S5 steps), it was found that only a small amount of ALB could be produced + / AFP + Liver precursor cells ( figure 2 a).

[0084] The present inventor finds after a large amount of research and experiment, because EPS cell has original state The characteristics of pluripotent stem cells, similar to the state of human embryonic pre-implantation epiblast 4 . The cells of the pre-implantation epiblast cannot directly respond to the developmental induction signals of the three germ layers, and the post-implantation epiblast cells have the ability to develop into cells of various lineages of the embryo 7. Therefore, we try to pretreat EPS cells first, induce them to the state of epiblast cells after implantation of early embryos, and then further dock the optimized differenti...

Embodiment 2

[0112] Example 2 Identification of differentiation efficiency and properties of each cell during the differentiation process of functional hepatic parenchymal cells

[0113] The inventors specifically analyzed the cells produced in the differentiation process:

[0114] First, we found that using TeSR TM 2 medium (prime stem cell medium) pretreats EPS cells, which can induce EPS cells to become epiblast cells after early embryo implantation. This stage is the first stage (S1), and the cells obtained in the S1 stage Named EPS-S1 cells.

[0115] In order to further study the state of EPS-S1 cells, the present invention uses the method of RNA sequencing (RNA-Sequencing) to analyze and compare the whole gene transcriptome of EPS-S1 cells, induced pluripotent stem cells (iPSCs) and embryonic stem cells (ESCs) , and human epiblast cells of different developmental stages were used as controls. It was found that EPS-S1 cells had a similar transcriptional profile to epithelial cells ...

Embodiment 3

[0123] Example 3 Properties and functions of functional hepatic parenchymal cells differentiated from EPS

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Abstract

The invention discloses functional liver parenchyma cells and a preparation method therefor. According to the preparation method, potential-extended multipotential stem cells, i.e., EPS cells are subjected to directed differentiation, so as to prepare the functional liver parenchyma cells (or called liver cells). Furthermore, before directed differentiation of the EPS cells, the EPS cells is firstly pretreated in an iPSCs / ESCs culture medium. According to the preparation method disclosed by the invention, human EPS cells can be more efficiently differentiated into the functional liver parenchyma cells, and the functional liver parenchyma cells EPS-Heps obtained through EPS differentiation are more similar to fresh-separated primary liver cells in a full-gene transcription level.

Description

technical field [0001] The invention belongs to the technical field of stem cell differentiation, and more specifically, the invention relates to a new method for inducing directional differentiation of pluripotent stem cells with expanded potential to functional liver parenchymal cells. Background technique [0002] The liver plays an important role in the metabolic process in the human body, including glycogen synthesis, urea synthesis, regulation of fat metabolism, and drug detoxification. Damage to liver cells, including hepatitis, liver cancer, and cirrhosis, will cause great harm to human life. To solve this problem, it is necessary to obtain a large number of liver cells in vitro and use these cells for transplantation. [0003] Among various cell sources, human pluripotent stem cells (pluripotent stem cells) have the ability of directed differentiation to produce a variety of functional cell types, which can be used in regenerative medicine and have great applicatio...

Claims

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Application Information

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IPC IPC(8): C12N5/071C12Q1/02
CPCC12N5/067G01N33/5067G01N33/5014C12N2501/16C12N2506/45C12N2500/38C12N2501/415C12N2501/117C12N2501/115C12N2501/155C12N2501/11C12N2500/30C12N2501/50C12N2501/15C12N2501/01C12N2501/405Y02A50/30
Inventor 邓宏魁王启明孙达时艳
Owner PEKING UNIV
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