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Simultaneous Maintenance of Human Cord Blood SCID-Repopulating Cells and Expansion of Committed Progenitors at Low O₂ Concentration (3%)

^a Laboratory for Studies on Hematopoiesis: Molecular and Functional Aspects, Bordeaux 2 University, Bordeaux, France;
^b Establishment Aquitaine-Limousin Regional Center, Bordeaux, France;
^c Laboratory of Hematology, Haut Lévêque Hospital, Pessac, France

Key Words. Severe combined immunodeficiency–repopulating cells • NOD/SCID Stem cells • IL-3 • Hypoxia • Cord blood • Ex vivo expansion

Correspondence: Zoran Ivanovic, M.D., Ph.D., Laboratoire Hématopoïèse Normale et Pathologique FRE CNRS 2617, Université Victor Segalen Bordeaux 2, Carreire Nord–Bât. 1B–RDC, 146, rue Léo Saignat–BP 50, 33076 Bordeaux Cedex, France. Telephone: 05-56-90-75-50; Fax: 05-56-90-75-51; e-mail: zoran.ivanovic{at}efs.sante.fr

In the present work, we tested the hypothesis that liquid cultures (LCs) of cord blood CD34⁺ cells at an appropriate low O₂ concentration could simultaneously allow colony-forming cell (CFC) expansion and nonobese diabetic/severe combined immunodeficiency mice–repopulating cell (SRC) maintenance. We first found that 3% was the minimal O₂ concentration, still allowing the same rate of CFC expansion as at 20% O₂. We report here that 7-day LCs of cord blood CD34⁺ cells at 3% O₂ maintain SRC better than at 20% O₂ and allow a similar amplification of CFCs (35- to 50-fold) without modifying the CD34⁺ cell proliferation. Their phenotypic profile (antigens: HLA-DR, CD117, CD33, CD13, CD11b, CD14, CD15, and CD38) was not modified, with exception of CD133, whose expression was lower at 3% O₂. These results suggest that low O₂ concentrations similar to those found in bone marrow participates in the regulation of hematopoiesis by favoring stem cell–renewing divisions. This expansion method that avoids stem cell exhaustion could be of paramount interest in hematopoietic transplantation by allowing the use of small-size grafts in adults.

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