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Efficient Generation and Editing of Feeder-free IPSCs from Human Pancreatic Cells Using the CRISPR-Cas9 System
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Bioengineering
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JoVE Journal Bioengineering
Efficient Generation and Editing of Feeder-free IPSCs from Human Pancreatic Cells Using the CRISPR-Cas9 System

Efficient Generation and Editing of Feeder-free IPSCs from Human Pancreatic Cells Using the CRISPR-Cas9 System

DOI:
10.3791/56260-v

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09:16 min

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November 08, 2017

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, ,
1Department of Animal and Avian Sciences,University of Maryland, 2Animal Bioscience and Biotechnology Laboratory, ARS,USDA, 3NIH Stem Cell Unit, Bethesda,National Institutes of Health, 4RenOVAte Biosciences Inc

Chapters

  • 00:05Title
  • 00:52Generation of Human iPSCs from Primary Human Pancreatic Cells
  • 03:41Genome Editing of iPSC Using CRISPER-Cas9
  • 05:45Single-cell Isolation of Targeted hiPSCs
  • 07:15Results: Editing of Feeder-free iPSCs with CRISPER-Cas9 System
  • 08:29Conclusion

Summary

Automatic Translation

Automatic Translation

This protocol describes in detail the generation of footprint-free induced pluripotent stem cells (iPSCs) from human pancreatic cells in feeder-free conditions, followed by editing using CRISPR/Cas9 ribonucleoproteins and characterization of the modified single-cell clones.

Tags

Keywords: Induced Pluripotent Stem CellsIPSCsCRISPR-Cas9Genome EditingFeeder-freeHuman Pancreatic CellsSendai VirusCell ReprogrammingCell CloningMEFsMTeSR1Matrix MembraneCell Culture

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