Journal
/
Genetics
/
Preparation and Gene Modification of Nonhuman Primate Hematopoietic Stem and Progenitor Cells
JoVE Journal
Genetics
A subscription to JoVE is required to view this content.  Sign in or start your free trial.
JoVE Journal Genetics
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

•

11:16 min

•

February 15, 2019

•

, , , , , , ,
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

Automatic Translation

Automatic Translation

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

Article

Embed

ADD TO PLAYLIST

Usage Statistics

Related Videos

Selection-dependent and Independent Generation of CRISPR/Cas9-mediated Gene Knockouts in Mammalian Cells

Sample Preparation and Analysis of RNASeq-based Gene Expression Data from Zebrafish

A Standard Methodology to Examine On-site Mutagenicity As a Function of Point Mutation Repair Catalyzed by CRISPR/Cas9 and SsODN in Human Cells

Immunostaining for DNA Modifications: Computational Analysis of Confocal Images

Generation of a Gene-disrupted Streptococcus mutans Strain Without Gene Cloning

Highly Efficient Gene Disruption of Murine and Human Hematopoietic Progenitor Cells by CRISPR/Cas9

Exploring the Root Microbiome: Extracting Bacterial Community Data from the Soil, Rhizosphere, and Root Endosphere

A Fast and Quantitative Method for Post-translational Modification and Variant Enabled Mapping of Peptides to Genomes

A Combinatorial Single-cell Approach to Characterize the Molecular and Immunophenotypic Heterogeneity of Human Stem and Progenitor Populations

Isolation, Characterization and MicroRNA-based Genetic Modification of Human Dental Follicle Stem Cells

Read Article