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Generation of Induced Pluripotent Stem Cells by Reprogramming Human Fibroblasts with the Stemgent Human TF Lentivirus Set
JoVE Journal
Biologia
This content is Free Access.
JoVE Journal Biologia
Generation of Induced Pluripotent Stem Cells by Reprogramming Human Fibroblasts with the Stemgent Human TF Lentivirus Set
DOI:

12:10 min

December 08, 2009

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Capítulos

  • 00:00Título
  • 00:09Introduction
  • 01:05Preparing Mouse Embryonic Fibroblast (MEF) Conditioned Medium
  • 02:39Transducing Human-Foreskin-Fibroblast (BJ) Cells with Stem-Cell-Specific Transcription Factors
  • 06:33Selecting and Passaging Induced Pluripotent Stem Cell Colonies
  • 07:33Staining Resulting Colonies for Markers of Pluripotency
  • 09:43Representative Results: Induced Pluripotent Stem Cell Colonies Morphologically Resemble Stem-Cells and Express Markers of Pluripotency
  • 11:07Conclusion

Summary

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We demonstrate the protocol for the generation of induced pluripotent stem cells from human somatic cells using lentivirus-mediated delivery of the human factors Oct4, Sox2, Nanog, and Lin28. Pluripotency was confirmed by morphology and the presence of embryonic stem (ES) cell-specific markers.

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