Very Low Oxygen Concentration (0.1%) Reveals Two FDCP-Mix Cell Subpopulations That Differ by Their Cell Cycling, Differentiation and P27KIP1 Expression

Oxygen (O(2)) concentrations in bone marrow vary from 4% in capillaries to <0.1% in subendosteum, in which hematopoietic stem cells reside in specific niches. Culture at low O(2) concentrations (3, 1 and 0.1%) influences hematopoietic stem and progenitor cells survival, proliferation and differentiation, depending on their level of differentiation. Culture of human CD34(+) cells at low O(2) concentrations (O(2) ≤3%) maintains stem cell engraftment potential better than at 20% O(2) (NOD/Scid xenograft model). In contrast, progenitors disappear from cultures at/or <1% O(2) concentrations. A very low O(2) concentration (0.1%) induces CD34(+) quiescence in G(0). The exploration of molecules and mechanisms involved in hematopoietic stem and progenitor cells' quiescence and differentiation related to low O(2) concentrations is unfeasible with primary CD34(+) cells. Therefore, we performed it using murine hematopoietic nonleukemic factor-dependent cell Paterson (FDCP)-Mix progenitor cell line. The culture of the FDCP-Mix line at 0.1% O(2) induced in parallel G(0) quiescence and granulo-monocytic differentiation of most cells, whereas a minority of undifferentiated self-renewing cells remained in active cell cycle. Hypoxia also induced hypophosphorylation of pRb and increased the expression of p27(KIP1), the two proteins that have a major role in the control of G(0) and G(1) to S-phase transition.