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Normal Hemopoiesis And Stemcellology

The expansion of murine bone marrow cells preincubated in hypoxia as an in vitro indicator of their marrow-repopulating ability

Leukemia volume 14, pages 735–739 (2000)Cite this article

Abstract

In liquid cultures of murine bone marrow cells stimulated with interleukin-3 and granulocyte/macrophage colony-stimulating factor, hypoxia (1% oxygen) induced a reversible block of hematopoiesis, maintaining the progenitors’ expansion potential unreduced. Progenitors repopulating day-14 hypoxic cultures with cells or granulocyte/macrophage colony-forming units (CFU-GM) were found, on the basis of their maintenance in hypoxia (12% and 76%, respectively), to belong to different subsets, the latter being much more efficiently maintained. The maintenance in hypoxic cultures of progenitors detectable by marrow-repopulating ability (MRA) assay was 18% for MRAcell progenitors and 69% for MRACFU progenitors. Thus, the repopulation of hypoxic cultures with cells or CFU-GM closely reflected the presence of progenitors capable of repopulating, with cells or CFU-GM, the bone marrow of lethally irradiated syngeneic animals. Progenitors repopulating hypoxic cultures were, like MRA progenitors, significantly resistant to 5- fluorouracil, progenitors repopulating cultures with CFU-GM being two-fold more resistant than those repopulating cultures with cells. We concluded that the repopulation of day-14 hypoxic cultures occurring after their transfer to air is to be considered an indicator of the maintenance of MRA progenitors in hypoxia. The relevance of these results to stem cell biology and their potential practical applications are discussed.

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Acknowledgements

The project was funded by grants from Ministero della Università e della Ricerca Scientifica e Tecnologica (funds 60% and 40%), Consiglio Nazionale delle Ricerche (CNR), Associazione Italiana per la Ricerca sul Cancro (AIRC) and Regione Toscana (Progetto Qualità). ZI was the recipient of a fellowship from AIRC. VP was supported by a grant from Conseil Regional du Limousin.

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  1. Dipartimento di Patologia e Oncologia Sperimentali, Università di Firenze, Firenze, Italy

    MG Cipolleschi, E Rovida, Z Ivanovic, M Olivotto & P Dello Sbarba

  2. Laboratoire d'Hématologie Expérimentale, Université de Limoges, Limoges, France

    V Praloran

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  1. MG Cipolleschi
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Cipolleschi, M., Rovida, E., Ivanovic, Z. et al. The expansion of murine bone marrow cells preincubated in hypoxia as an in vitro indicator of their marrow-repopulating ability. Leukemia 14, 735–739 (2000). https://doi.org/10.1038/sj.leu.2401744

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