A novel strategy to derive iPS cells from porcine fibroblasts |
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Authors: | WeiMin Ruan JianYong Han Pin Li SuYing Cao Yang An Bing Lim Ning Li |
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Institution: | (1) Center for iPS cell Research and Application, Institute for Integrated Cell-Material Sciences, Kyoto University, Kyoto 606-8507, Japan;(2) Department of Stem Cell Biology, Institute for Frontier Medical Sciences, Kyoto University, Kyoto 606-8507, Japan;(3) Yamanaka iPS Cell Special Project, Japan Science and Technology Agency, Kawaguchi 332-0012, Japan;(4) Gladstone Institute of Cardiovascular Disease, San Francisco, CA 94158, USA |
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Abstract: | Induced pluripotent stem (iPS) cell technology demonstrates that somatic cells can be reprogrammed to a pluripotent state
by over-expressing four reprogramming factors. This technology has created an interest in deriving iPS cells from domesticated
animals such as pigs, sheep and cattle. Moloney murine leukemia retrovirus vectors have been widely used to generate and study
mouse iPS cells. However, this retrovirus system infects only mouse and rat cells, which limits its use in establishing iPS
cells from other mammals. In our study, we demonstrate a novel retrovirus strategy to efficiently generate porcine iPS cells
from embryonic fibroblasts. We transfected four human reprogramming factors (Oct4, Sox2, Klf4 and Myc) into fibroblasts in one step by using a VSV-G envelope-coated pantropic retrovirus that was easily packaged by GP2-293 cells.
We established six embryonic stem (ES)-like cell lines in human ES cell medium supplemented with bFGF. Colonies showed a similar
morphology to human ES cells with a high nuclei-cytoplasm ratio and phase-bright flat colonies. Porcine iPS cells could form
embryoid bodies in vitro and differentiate into the three germ layers in vivo by forming teratomas in immunodeficient mice. |
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