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In vitro induction of embryo stem cell source elliptocyte and separation purification culturing method

A technology of embryonic stem cells and stem cells, which is applied in the field of in vitro induction, isolation, purification and culture of embryonic stem cell-derived hepatic oval cells, which can solve the problems of cumbersome and time-consuming induction model induction process

Inactive Publication Date: 2007-04-18
JINAN UNIVERSITY
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Problems solved by technology

[0005] The technical problem to be solved by the present invention is: to solve the defects in the prior art that the induction process of the rodent oval cell induction model is cumbersome and time-consuming, and the inducers are all chemical toxic substances, which have adverse effects, thus limiting the scope of its application; In the process of directional induction of embryonic stem cells in vitro to generate hepatic parenchymal cells, there is an intermediate differentiation stage of embryonic stem cell-derived hepatic oval cells with bidirectional differentiation ability, and HGF and EGF have a significant effect on the generation of embryonic stem cell-derived hepatic oval cells induction; and rapid and convenient induction and sorting of suitable hepatic oval cells

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  • In vitro induction of embryo stem cell source elliptocyte and separation purification culturing method
  • In vitro induction of embryo stem cell source elliptocyte and separation purification culturing method
  • In vitro induction of embryo stem cell source elliptocyte and separation purification culturing method

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Embodiment Construction

[0020] The following specific examples are only to further describe the technical solutions, and do not limit the scope of the present invention.

[0021] (1) Experimental materials and equipment:

[0022] 1. Source of embryonic stem cells:

[0023] Commercially available Balb / c mouse embryonic stem cells were selected.

[0024] 2. Cell culture reagents:

[0025] 2.1 Cell culture medium:

[0026] (1) Components of Embryonic Stem Cell Medium:

[0027] DMEM (Dubecco's modified Eagle'smedium) containing high sugar and glutamine (Hyclone, USA);

[0028] 20% fetal bovine serum (Fetal bovine serum, FBS) (Hangzhou Sijiqing Company);

[0029] 1000U / ml recombinant mouse leukemia inhibitory factor (recombinant murineleukemia inhibitory factor, LIF) (USA ESGRO  -Chemicon Corporation);

[0030] 0.1mmol / L β-mercaptoethanol (β-Mercaptoethanol, β-ME) (Invitrogen, USA);

[0031] 25mmol / L HEPES (Invitrogen, USA);

[0032] 100 U / ml penicillin and 100 μg / ml streptomycin (Invitrogen, US...

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Abstract

This invention provides a efficiently new induction model of hepatic oval cell. In this model significantly inducing the production of embryonic stem cell endogenous oval cell by using HGF and EGF, the whole induction method is fairly convenient, induction cycle is also very short. Utilizing very little embryonic stem cell provenance source can induce to generate sufficient hepatic oval cell which is contented to clinical requirement by using the invented induction model of hepatic oval cell, it also can be used for clinical category such as hepatic oval cell transplant, hepatic tissue engineering and so on, it can play a part in assistant supportive treatment or curing disease for Liver function failure, hepatic metabolic disease caused by various kinds of reason.

Description

1. Technical field [0001] The invention relates to a method for in vitro induction, separation, purification and culture of cells, in particular to a method for in vitro induction, separation and purification of embryonic stem cell-derived hepatic oval cells. 2. Background technology [0002] Embryonic stem cells (ESCs) are a type of undifferentiated diploid cells isolated from the inner cell mass of early mammalian blastocysts, which have the potential of self-renewal and totipotent differentiation [1] . Under certain conditions, embryonic stem cells can maintain an undifferentiated state and can expand infinitely. They have the potential to differentiate into all tissues and cells in the body. They are an unlimited source of cells. The potential is huge. It is the dream of many researchers to control the directional differentiation of embryonic stem cells to generate various desired target cells for cell therapy or even to construct desired organs. The establishment of ...

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Application Information

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IPC IPC(8): C12N5/06C12N5/071C12N5/0735
Inventor 银东智蔡继业郑启昌刘美莉
Owner JINAN UNIVERSITY
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