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Ex vivo Expansion und genetische Manipulation von hämatopoetischen Stammzellen der Maus in Kulturen auf Polyvinylalkoholbasis
JoVE Journal
Biologia
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JoVE Journal Biologia
Ex Vivo Expansion and Genetic Manipulation of Mouse Hematopoietic Stem Cells in Polyvinyl Alcohol-Based Cultures
DOI:

07:55 min

February 10, 2023

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Capítulos

  • 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

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Hier wird ein Protokoll zur Initiierung, Aufrechterhaltung und Analyse hämatopoetischer Stammzellkulturen der Maus unter Verwendung von ex vivo Polyvinylalkohol-basierter Expansion sowie Methoden zur genetischen Manipulation dieser Zellen durch lentivirale Transduktion und Elektroporation vorgestellt.

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