Bone Marrow Extraction and c-Kit+ Enrichment of Hematopoietic Stem Cell (HSC)
3:03
Initiating Cell Cultures with c-Kit-Enriched Hematopoietic Stem and Progenitor Cells (HSPCs)
3:45
Performing Media Changes
4:19
Electroporating Cultured HSPCs
5:24
Transducing Cultured HSPCs with Lentiviral Vector
6:17
Results: Fluorescence-Activated Cell Sorting (FACS) of the c-Kit-Enriched HSCs
7:22
Conclusion
Transcription
This protocol permits the expansion and genetic manipulation of functional mouse hematopoietic stem cells, allowing a deeper understanding of hematopoietic stem cell biology and hematopoiesis. The advantage of this technique is that it can support
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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.