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Biologia
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Generation of Induced Pluripotent Stem Cells by Reprogramming Mouse Embryonic Fibroblasts with a Four Transcription Factor, Doxycycline Inducible Lentiviral Transduction System
JoVE Journal
Biologia
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JoVE Journal Biologia
Generation of Induced Pluripotent Stem Cells by Reprogramming Mouse Embryonic Fibroblasts with a Four Transcription Factor, Doxycycline Inducible Lentiviral Transduction System

Generation of Induced Pluripotent Stem Cells by Reprogramming Mouse Embryonic Fibroblasts with a Four Transcription Factor, Doxycycline Inducible Lentiviral Transduction System

DOI:
10.3791/1447-v

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11:48 min

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November 13, 2009

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, , ,
1Stemgent, 2Whitehead Institute for Biomedical Research,MIT – Massachusetts Institute of Technology

Capítulos

  • 00:00Título
  • 00:09Introduction
  • 00:30Viral Transduction of Mouse Embryonic Fibroblasts
  • 01:25Doxycycline Induced Reprogramming
  • 03:18Immunocytochemical Analysis of Transduction Efficiency
  • 05:31Isolating and Expanding Induced Pluripotent Stem Cell Colonies
  • 07:39Immunocytochemical Analysis for Pluripotency
  • 09:14Pluripotency Marker Analysis
  • 10:31Conclusion

Summary

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The Stemgent Dox Inducible Mouse TF Lentivirus Set can reprogram mouse embryonic fibroblasts (MEFs) to induced pluripotent stem (iPS) cells. Here we demonstrate the protocol for DOX-inducible expression of mouse reprogramming transcription factors Oct4, Sox2, Klf4 and c-Myc to generate iPS colonies that express common mES pluripotency markers.

Tags

Induced Pluripotent Stem CellsReprogrammingTranscription FactorsDoxycycline Inducible Lentiviral Transduction SystemMouse Embryonic FibroblastsPluripotencyRetrovirus-mediated DeliveryOct4Sox2C-MycKlf4Doxycycline Inducible Lentiviral Delivery SystemAdult Mouse Somatic Cell TypesEmbryonic Stem CellsRegenerative MedicineTeratoma FormationDifferentiated CellsEthical DilemmaStemgent DOX Inducible Mouse TF Lentivirus Set

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