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Characterization of retroviral gene transfer into highly purified human CD34(-) cells with primitive hematopoietic capacity. | LitMetric

AI Article Synopsis

  • Recent research has identified a specific group of primitive human hematopoietic cells, specifically CD34(-) AC133(+) Lin(-) cells, which show a higher efficiency for retroviral transduction compared to other subpopulations.
  • These CD34(-) AC133(+) Lin(-) cells not only expanded during culture but also demonstrated successful gene transfer, evidenced by the expression of enhanced green fluorescent protein (eGFP).
  • This study is the first of its kind to evaluate retroviral transduction in these primitive cells, paving the way for improving gene transfer techniques for clinical applications involving CD34(-) stem cells.

Article Abstract

Primitive human hematopoietic cells have recently been identified within a rare subfraction of CD34(-) lineage-depleted (Lin(-)) cells and further characterized by their restriction to a rarer subset expressing AC133. Here we show that CD34(-)AC133(+)Lin(-) cells can be transduced by retrovirus at a comparatively higher efficiency than either CD34(-)AC133(-)Lin(-) or CD34(+)CD38(-)Lin(-) cells. Subpopulations were transduced by enhanced green fluorescent protein (eGFP)-containing retrovirus in serum-free conditions. During the culture period, both CD34(-)AC133(+)Lin(-) and CD34(+)CD38(-)Lin(-) subfractions expanded, whereas CD34(-)AC133(-)Lin(-) cells could not be sustained. Fluorescent microscopic examination of progenitors assayed by colony-forming units (CFU) derived from CD34(-)AC133(+)Lin(-) cells revealed expression of eGFP, with the presence of provirus confirmed by clonal PCR analysis. Flow cytometry detecting eGFP revealed that cultures seeded with CD34(-)AC133(+)Lin(-) cells had a greater than threefold higher frequency of eGFP(+) cells compared with transduced cultures of CD34(+)CD38(-)Lin(-) cells. Our results demonstrate that retroviral transduction efficiency and level of transgene expression into CD34(-)AC133(+)Lin(-) cells is distinct to either CD34(-)AC133(-)Lin(-) or CD34(+)CD38(-)Lin(-) cells. This study represents the first evaluation of retroviral transduction into this population of primitive CD34(-) cells, and therefore provides the basis for optimization of gene transfer protocols to examine the role of gene-marked CD34(-) stem cells in a clinical setting.

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Source
http://dx.doi.org/10.1006/mthe.2002.0583DOI Listing

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