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Ex Vivo Expansion and Genetic Manipulation of Mouse Hematopoietic Stem Cells in Polyvinyl Alcohol-Based Cultures
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Ex Vivo Expansion and Genetic Manipulation of Mouse Hematopoietic Stem Cells in Polyvinyl Alcohol-Based Cultures

Ex Vivo Expansion and Genetic Manipulation of Mouse Hematopoietic Stem Cells in Polyvinyl Alcohol-Based Cultures

DOI:
10.3791/64791-v

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07:55 min

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February 10, 2023

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, ,
1MRC Weatherall Institute of Molecular Medicine,University of Oxford

Chapters

  • 00:05Introduction
  • 01:05Bone Marrow Extraction and c-Kit&#43 Enrichment of Hematopoietic Stem Cell &#40HSC&#41
  • 03:03Initiating Cell Cultures with c-Kit-Enriched Hematopoietic Stem and Progenitor Cells &#40HSPCs&#41
  • 03:45Performing Media Changes
  • 04:19Electroporating Cultured HSPCs
  • 05:24Transducing Cultured HSPCs with Lentiviral Vector
  • 06:17Results: Fluorescence-Activated Cell Sorting &#40FACS&#41 of the c-Kit-Enriched HSCs
  • 07:22Conclusion

Summary

Automatic Translation

Automatic Translation

Presented here is a protocol to initiate, maintain, and analyze mouse hematopoietic stem cell cultures using ex vivo polyvinyl alcohol-based expansion, as well as methods to genetically manipulate them by lentiviral transduction and electroporation.

Tags

Keywords: Hematopoietic Stem CellsEx Vivo ExpansionGenetic ManipulationPolyvinyl AlcoholMouseBone MarrowC-KitMagnetic Column SeparationCell Culture

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