Background:Mesenchymal stem cells derived from bone marrow and adipose tissue are being applied to tissue engineering and cell therapy. The use of stem cells of various clinical applications is highly expected and the production of good quality stem cells is very critical for basic studies. In the bone marrowmesenchymal stem cells from an unique niche in which the oxygen tension is low. Hypoxia may have a role in maintaining stem cell fate, self renewal and multi-potency. We investigated whether low oxygen culture would be beneficial for mesenchymal stem cell. Results:BMCs from 8-10 week aged, 6 mice were subjected to hypoxic conditioning by culture for 7 days in 20%, 3%, 1% oxygen. For culture 1x106 cell/ml were seeded in media with L-glutamine in each dish. During the culturing, cell colonies were checked once in three days. After cell culture, we stained cells by CD90 then counted CD90 positive cells by fluorescence microscope. More colonies and mesenchymal cells were observed at 3%, 1% oxygen and also colonies were bigger in hypoxic condition. Statistical significances were identified mesenchymal cells (p<0.05) in hypoxic condition. Conclusions:Our data suggests low physiological oxygen culture could improve the differentiation of mesenchymal cells. Long term culture will be necessary to confirm whether low physiological oxygen levels also improve genomic stability.