Introduction The identification of potential breast cancer stem cells is of importance as the characteristics of stem cells suggest that they are resistant to conventional forms of therapy. Several techniques have been proposed to isolate or enrich for tumorigenic breast cancer stem cells, including (a) culture of cells in non-adherent non-differentiating conditions to form mammospheres and (b) sorting of the cells by their surface phenotype (expression of CD24 and CD44).

Articular cartilage is a physiologically hypoxic tissue with a proposed gradient of oxygen tension ranging from about 10% oxygen at the cartilage surface to less than 1% in the deepest layers. The position of the chondrocyte within this gradient may modulate the cell's behavior and phenotype. Moreover, the oxygen gradient is likely to be disturbed during joint diseases in which the pO(2)of the synovial fluid declines which may cause changes in chondrocyte behavior and gene expression. Thus, there is a need to understand the chondrocyte's response to different oxygen tensions. We compared the behavior of bovine articular chondrocytes cultured in alginate beads for 7 days in medium maintained at <0.1, 5, 10 or 20% oxygen. The chondrocytes' survival, differentiation, cell division, viability and matrix production were assessed at each oxygen tension and rRNA and mRNA abundance was measured. Chondrocytes were able to survive under all oxygen tensions for at least 7 days but cells cultured under anoxic conditions were metabolically less active than cells maintained in higher oxygen tensions; this was associated with a decrease in matrix production. In <0.1% oxygen there was a marked decrease in rRNA and mRNA abundance in the cells. There were no differences in cell division or differentiation between any oxygen tensions. These findings indicate that articular chondrocytes can be cultured successfully in the pO(2)range in which they are thought to exist in vivo (5-10% pO(2)) and are fully active under these conditions. Under anoxic conditions (<0.1% pO(2)) function is severely compromised.