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Selecting and Isolating Colonies of Human Induced Pluripotent Stem Cells Reprogrammed from Adult Fibroblasts
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生物学
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JoVE 杂志 生物学
Selecting and Isolating Colonies of Human Induced Pluripotent Stem Cells Reprogrammed from Adult Fibroblasts

Selecting and Isolating Colonies of Human Induced Pluripotent Stem Cells Reprogrammed from Adult Fibroblasts

DOI:
10.3791/3416-v

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13:23 min

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February 20, 2012

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, , , , ,
1Department of Molecular Carcinogenesis and Center for Cancer Epigenetics,University of Texas M.D. Anderson Cancer Center, 2Department of Cell Biology,Poznan University of Medical Sciences, 3Department of Molecular Biology,The Scripps Research Institute

Chapters

  • 00:05Title
  • 01:31Virus Production
  • 03:53Viral Transduction
  • 05:47Reprogramming
  • 07:24Isolation of Human iPS Cell Colonies
  • 11:14Representative Results: Examining Transduction Efficiency and Characterizing Reprogrammed Cells
  • 12:58Conclusion

Summary

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We present a protocol for efficient reprogramming of human somatic cells into human induced pluripotent stem cells (hiPSC) using retroviral vectors encoding Oct3/4, Sox2, Klf4 and c-myc (OSKM) and identification of correctly reprogrammed hiPSC by live staining with Tra-1-81 antibody.

Tags

SelectingIsolatingColoniesHuman Induced Pluripotent Stem CellsReprogrammingAdult FibroblastsRetroviral VectorsOct3/4Sox2Klf4C-mycSodium ButyrateEfficient Transduction ProtocolVirus-containing MediaLive ImmunostainingTra-1-81Pluripotent CellsManually PassagedMatrigel PlatesFeeder-free ConditionsHES-like MorphologyExpansionCell LinesPluripotency MarkersTRA-1-60SSEA-4Oct3/4Sox2Nanog

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