Efficient iPS Cell Production with the MyoD Transactivation Domain in Serum-Free Culture

Autor: Peter Karian, Meri T. Firpo, Nobuko Katoku-Kikyo, Hiroyuki Hirai, Nobuaki Kikyo
Jazyk: angličtina
Rok vydání: 2012
Předmět:
Cellular differentiation
Cell Culture Techniques
lcsh:Medicine
Gene Expression
MyoD
Culture Media
Serum-Free
Myoblasts
Transactivation
Mice
0302 clinical medicine
lcsh:Science
Induced pluripotent stem cell
0303 health sciences
Multidisciplinary
Stem Cells
Teratoma
Genomics
KLF4
030220 oncology & carcinogenesis
Epigenetics
Genetic Engineering
Reprogramming
Research Article
Biotechnology
Transcriptional Activation
Induced Pluripotent Stem Cells
Mice
Transgenic
Biology
Molecular Genetics
03 medical and health sciences
Kruppel-Like Factor 4
SOX2
Genetics
Animals
Humans
030304 developmental biology
MyoD Protein
lcsh:R
Fibroblasts
Alkaline Phosphatase
Chromatin Assembly and Disassembly
Embryonic stem cell
Molecular biology
Coculture Techniques
Protein Structure
Tertiary
Microscopy
Fluorescence
lcsh:Q
Octamer Transcription Factor-3
Developmental Biology
Zdroj: PLoS ONE
PLoS ONE, Vol 7, Iss 3, p e34149 (2012)
ISSN: 1932-6203
Popis: A major difficulty of producing induced pluripotent stem cells (iPSCs) has been the low efficiency of reprogramming differentiated cells into pluripotent cells. We previously showed that 5% of mouse embryonic fibroblasts (MEFs) were reprogrammed into iPSCs when they were transduced with a fusion gene composed of Oct4 and the transactivation domain of MyoD (called M(3)O), along with Sox2, Klf4 and c-Myc (SKM). In addition, M(3)O facilitated chromatin remodeling of pluripotency genes in the majority of transduced MEFs, including cells that did not become iPSCs. These observations suggested the possibility that more than 5% of cells had acquired the ability to become iPSCs given more favorable culture conditions. Here, we raised the efficiency of making mouse iPSCs with M(3)O-SKM to 26% by culturing transduced cells at low density in serum-free culture medium. In contrast, the efficiency increased from 0.1% to only 2% with the combination of wild-type Oct4 and SKM (OSKM) under the same culture condition. For human iPSCs, M(3)O-SKM achieved 7% efficiency under a similar serum-free culture condition, in comparison to 1% efficiency with OSKM. This study highlights the power of combining the transactivation domain of MyoD with a favorable culture environment.
Databáze: OpenAIRE
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