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TISSUE-SPECIFIC STEM CELLS

Very Low O₂ Concentration (0.1%) Favors G₀ Return of Dividing CD34⁺ Cells

^a CNRS UMR 5164: University of Bordeaux 2, Bordeaux, France;
^b University Hospital, Hematology Laboratory, Bordeaux, France;
^c French Blood Transfusion Organisation in Aquitaine-Limousin, Bordeaux, France

Key Words. CD34 • Cell cycle • Hematopoietic stem cell • Hypoxia • Oxygen

Correspondence: Vincent Praloran, M.D., Ph.D., CNRS UMR 5164, Université de Bordeaux 2, 33076 Bordeaux CEDEX, France. Telephone: 33-5-57–57-16-11; Fax: 33-5-56-51-42-18; e-mail: Vincent.Praloran{at}hemato.u-bordeaux2.fr

Physiological bone marrow oxygen concentrations are everywhere lower than 4% and almost null in some areas. We compared the effects of 20%, 3%, and 0.1% O₂ concentrations on cord blood CD34⁺ cell survival, cycle, and functionality in serum-free cultures for 72 hours with or without interleukin-3 (IL-3). As from 24 hours, IL-3 improved cell survival and proliferation in all conditions. After 72 hours, cells were 1.5 and 2.5 times more in quiescence (G₀) at 3% and 0.1% O₂, respectively, than at 20%; transforming growth factor-ß signaling seemed not to be involved. To explore cell cycle further, fresh CD34⁺ cells were stained with PKH26 and cultured for 72 hours, and then undivided and divided cells were sorted. At 0.1% O₂, 46.5% ± 19.1% of divided cells returned to G₀ compared with 7.9% ± 0.3% at 20%. Colony formation and nonobese diabetic/severe combined immunodeficient mice engraftment efficiency were similar after 3 days at 20% and 0.1% O₂ concentrations but lower than at T₀. In conclusion, a low O₂ concentration, close to those found in bone marrow stem cell niches, induces the G₀ return of CD34⁺ cells without impairing their functional capacity.

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