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Selektion og isolering Kolonier af Human induceret pluripotente stamceller omprogrammeret fra voksne fibroblaster
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JoVE Journal Biology
Selecting and Isolating Colonies of Human Induced Pluripotent Stem Cells Reprogrammed from Adult Fibroblasts

Selektion og isolering Kolonier af Human induceret pluripotente stamceller omprogrammeret fra voksne fibroblaster

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

Automatic Translation

Automatic Translation

Vi præsenterer en protokol til effektiv omprogrammering af humane somatiske celler i menneskelige inducerede pluripotente stamceller (hiPSC) ved hjælp af retrovirale vektorer koder Oct3 / 4, Sox2, Klf4 og c-myc (OSKM) og identifikation af korrekt omprogrammeres hiPSC af levende farvning med Tra- 1-81 antistof.

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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