Culture system for preparing endoderm cell line and method for preparing endoderm cell line

Abstract

The invention provides a culture system for preparing an endoderm cell line and a method for preparing the endoderm cell line by the culture system. The culture system comprises a composition of a TGF-beta receptor antagonist, an RA receptor activator and a cAMP (cyclic adenosine monophosphate) activating agent, wherein the composition is free from exogenous transcription factors. By the aid of the culture system and the culture method, reprogramming for changing cell fate can be induced, and body cells are reprogrammed into the endoderm cell line.

Description

technical field [0001] The invention relates to a culture system for preparing endoderm cell lines by inducing the reprogramming of small chemical molecules and growth factors to change cell fate, and a method, kit and application for preparing endoderm cell lines using the culture system. Background technique [0002] All organisms rely on transcriptional machinery to express specific portions of the genome in response to changes in the environment or developmental signals, thereby performing key biological functions throughout the life cycle. Thus, the transcription process constitutes a critical step in the regulation of biological processes, and transcription factors are considered to be master switches in determining cell fate. In the past decade, the rapid development of stem cell biology has mainly benefited from the elucidation of the functions of several transcription factors, which are the main regulators of the pluripotency of induced pluripotent stem cells (iPSCs...

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Problems solved by technology

[0003] However, although the research progress of transcription factor-based reprogramming to change cell fate is of great significance to the research of regenerative medicine and life sciences, overexpression of exogenous transcription factors through retrovirus integration into the genome of somatic cells There are potential safety issues, such as the use of the proto-oncogene C-Myc leading to tumor formation (Nakagawa et al., 2007), which hinder the clinical application of this technology (Hawley, 2008, Does retroviral insertional mutagenesis play a role in the generation of induced pluripotent stem cells? Molecular therapy: the journal of the American Society of GeneTherapy 16, 1354-1355; Lister et al., 2011, Hotspots of aberrant epigenomic reprogramming in human induced pluripotent stem cells. al., 2008, Dissecting direct reprogramming through integrative genomic analysis. Nature 454, 49-55; Ohi et al., 2011, Incomplete DNA methylation underlies a transcriptional memory of somatic cells in human iPS cells. Nature Cell Biology 13, 541-549 , 2007, Generation of germline-competent induced pluripotent stem cells. Nature 448, 313-317)

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