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Preparation and Gene Modification of Nonhuman Primate Hematopoietic Stem and Progenitor Cells
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Preparation and Gene Modification of Nonhuman Primate Hematopoietic Stem and Progenitor Cells

Preparation and Gene Modification of Nonhuman Primate Hematopoietic Stem and Progenitor Cells

DOI:
10.3791/58933-v

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11:16 min

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February 15, 2019

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, , , , , , ,
1Stem Cell and Gene Therapy Program,Fred Hutchinson Cancer Research Center, 2Department of Medicine,University of Washington, 3Department of Pathology,University of Washington

Chapters

  • 00:04Title
  • 00:42Bone Marrow Hemolysis (Day -1)
  • 02:44CD34-enriched Cell Quality Control (Day -1)
  • 05:10CD34+ Hematopoietic Stem and Progenitor Cell (HSPC) Gene Modification and Overnight Recovery (Day 0)
  • 06:33Cell Harvest and Infusion Preparation (Day 1)
  • 07:42Liquid Culture Quality Control (Days 1-5)
  • 09:19Results: Representative HSPC Identification and Analysis
  • 10:34Conclusion

Summary

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The goal of this protocol is to isolate nonhuman primate CD34+ cells from primed bone marrow, to gene-modify these cells with lentiviral vectors, and to prepare a product for infusion into the autologous host. The total protocol length is approximately 48 h.

Tags

Nonhuman PrimateHematopoietic Stem CellsProgenitor CellsGene TherapyBone MarrowCell IsolationCD34Magnetic Cell SortingFlow CytometryCell Culture

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