What question did this study set out to answer?

To develop a retroviral vector encoding murine heat-stable antigen to enable the rapid, non-toxic identification and physical isolation of transduced human hematopoietic progenitor and stem cells.

January 15, 1996Blood

Rapid and efficient selection of human hematopoietic cells expressing murine heat-stable antigen as an indicator of retroviral-mediated gene transfer

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Authors

ECEibhlin ConneallyUniversity of British Columbia Hospital PBPeter BardyRoyal Adelaide Hospital CECJ EavesBC Cancer Agency

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Implication

In vitro study demonstrates rapid marker selection of gene-modified human hematopoietic cells, indicating an efficient strategy for non-toxic enrichment in gene therapy.

Key Points

To develop a retroviral vector encoding murine heat-stable antigen to enable the rapid, non-toxic identification and physical isolation of transduced human hematopoietic progenitor and stem cells.
Constructed a murine stem cell virus (MSCV) vector encoding murine heat-stable antigen (HSA) and packaged it in helper-free amphotropic AM12 producer cells.
Cocultured cytokine-stimulated human bone marrow or mobilized peripheral blood CD34+ cells with producer cells for 48 hours.
Evaluated gene transfer and sorting efficacy 24 hours post-infection using fluorescence-activated cell sorting (FACS) and G418-resistant colony-forming progenitor assays across 4 to 6 experiments.
FACS analysis 24 hours post-transduction revealed that 21% to 41% (mean, 27%) of CD34+ cells expressed HSA, extending to primitive CD34+CD38- subpopulations.
FACS isolation of CD34+HSA+ cells increased gene transfer efficiency from 11% to 100% in erythroid progenitors and from 12% to 77% in granulopoietic progenitors, compared to 2% and 5% in CD34+HSA- fractions.
Gene transfer to long-term culture-initiating cells rose from ≤7% in unsorted cells to 86% in the CD34+HSA+ fraction, while dropping to 3% in CD34+HSA- cells.

Cite This Study

Conneally et al. (1996) studied this question.

synapsesocial.com/papers/6a8ce0eafe7e19d65ef55323 https://doi.org/10.1182/blood.v87.2.456.bloodjournal872456

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Implication

In vitro study demonstrates rapid marker selection of gene-modified human hematopoietic cells, indicating an efficient strategy for non-toxic enrichment in gene therapy.

Rapid and efficient selection of human hematopoietic cells expressing murine heat-stable antigen as an indicator of retroviral-mediated gene transfer

Discussion

Member takes

Implication

Key Points

Cite This Study

Rapid and efficient selection of human hematopoietic cells expressing murine heat-stable antigen as an indicator of retroviral-mediated gene transfer

Discussion

Member takes

Implication

Key Points

Cite This Study

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