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Exp Hematol. 1995 Aug;23(9):1011-5.

Long-term repopulating abilities of enriched fetal liver stem cells measured by competitive repopulation.

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  • 1Department of Molecular Biology, Princeton University, NJ, USA.

Abstract

To characterize hematopoietic cell biology, many investigators have used protocols that enrich for primitive hematopoietic stem cells (PHSC). In this study, we quantified the long-term repopulating ability (LTRA) of enriched and discarded fractions of PHSC from day-14 murine fetal liver using the competitive repopulation assay. We fractionated populations of fetal cells using the antigenic markers AA4.1+, AA4.1+/Sca+, and AA4.1+/Linlow/Sca+. Differentiating and repopulating abilities of each of these populations were directly compared using competitive repopulation. Adult bone marrow was mixed with fetal cell fractions from congenic donors having genetically distinguishable markers, and mixtures were given to irradiated recipients. Differentiating and repopulating abilities of the enriched donor cells were measured by the proportions of myeloid and lymphoid cells having donor markers that repopulated the recipients. LTRA was found primarily in the AA4.1+ and AA4.1+/Sca+ subpopulations. Further fractionation of the AA4.1+ cells to derive an AA4.1+/Linlow/Sca+ fraction showed that virtually all of the long-term stem cell activity was found in this subpopulation. These cells were 1400- to 1600-fold enriched in long-term functional ability compared to fresh marrow. This very high multilineage repopulating ability per cell was directly measured using a long-term functional assay in vivo. Importantly, the measured repopulating ability for AA4.1+/Linlow/Sca+ cells was about five-fold less than expected from the fraction of cells enriched and remained two- to three-fold less even after compensating for repopulating ability in discarded fractions. This illustrates that long-term functional abilities of enriched PHSC cannot be estimated from fractions enriched but should be quantitatively assayed.

PMID:
7635180
[PubMed - indexed for MEDLINE]
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