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Vitro hepatic differentiation

A Liver Differentiation, Population Technology

Active Publication Date: 2013-06-26
CAMBRIDGE ENTERPRISE LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Such studies are limited by the difficulty of culturing primary hepatocytes and the inability to provide relevant human stem cell-like cell lines that faithfully replicate the disease-causing protein dysfunction and consequent cellular defects [12]

Method used

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  • Vitro hepatic differentiation
  • Vitro hepatic differentiation
  • Vitro hepatic differentiation

Examples

Experimental program
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Embodiment Construction

[0189] experiment

[0190] method

[0191] hIPSC derivation and culture

[0192] After appropriate ethical review and patient consent, 8 mm skin punch biopsies were obtained from volunteer patients attending Addenbrooke's Hospital (Ethics Committee No. 08 / H0311 / 201; R&D No: A091485). Fibroblasts were obtained from tissue donations under GMP conditions using standardized laboratory protocols and expanded in standard fibroblast medium. Additional fibroblast samples were obtained from INSERM (France) and Coriell Biorepository. As detailed in Table 1, a total of 5 different disease samples were obtained from 7 different patients. Moloney murine leukemia virus-derived vectors each contained the coding sequence for one of four human genes (Oct-4, Sox2, c-Myc, and Klf4), while the corresponding viral particles were produced by Vectalys (Toulouse, France). ) were generated and used to infect fibroblasts with a 10-fold multiplicity of infectivity, as originally described by Y...

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Abstract

This invention relates to the induction of hepatic differentiation by culturing induced pluripotent stem (iPS) cells in an endoderm induction medium to produce a population of anterior definitive endoderm (ADE) cells and then culturing the population of ADE cells in a hepatic induction medium to produce a population of hepatic progenitor cells, which may be optionally differentiated into hepatocytes. The endoderm induction medium is a chemically defined medium which has fibroblast growth factor activity, stimulates SMAD2 and SMAD3 mediated signalling pathways and SMAD1, SMAD5 and SMAD9 mediated signalling pathways, and inhibits phosphatidylinositol 3-kinase (PI3K) and glycogen synthase kinase 3beta (GSK3beta); and the hepatic induction medium is a chemically defined medium which stimulates SMAD2 and SMAD3 mediated signalling pathways. These methods may be useful, for example, in producing hepatocytes and hepatic progenitor cells for cell-based therapies or disease modelling.

Description

technical field [0001] The present invention relates to the in vitro induction of hepatic differentiation in pluripotent human cells, especially human iPS cells. Background technique [0002] The possibility to derive human induced pluripotent stem cells (hIPSCs) by overexpressing some transcription factors in somatic cells opens up new opportunities for regenerative medicine and in vitro disease modeling [1]. hIPSCs have been generated from patients with various diseases [2][3][4], and when these cells are subsequently differentiated into neural progenitor cells they have multiple sets of reporter disease-specific phenotypes [5][6]. However, to date, no hIPSC-based models have been reported for use in non-neuronal cells, such as cells of mesoderm and endoderm lineage, nor as a function observed only in fully differentiated adult cells. Diseases that occur as a result of loss (late-onset disease). Furthermore, there remains concern that cellular stresses inherent in reprog...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071
CPCC12N5/067C12N2501/115C12N2501/155C12N2501/16C12N15/85
Inventor 卢多维克·瓦利耶谢赫·塔米尔·拉希德尼古拉斯·汉南卓欣桦
Owner CAMBRIDGE ENTERPRISE LTD
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