1.RESULTS OF CULTURING STEM CELLS DERIVED FROM MOUSE BONE MARROW UNDER VARYING OXYGEN CONDITIONS
Bayarmaa E ; Hayashi Humiko ; Byambasuren D ; Bayarjavkhlan Ch ; Naran G ; Tao-Sheng Li
Innovation 2017;11(3):14-18
BACKGROUND: In the recent years, mesenchymal stem cells have become increasingly utilized in regenerative medicine and tissue engineering applications because of their properties for self-renewal, differentiation and immunoregulation. 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 marrow, hematopoietic and mesenchymal 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 hematopoietic stem cell and mesenchymalstemcell.
MATERIAL: BMCs from 8-12 week aged, 15 mice were subjected to hypoxic conditioning by culture for 8-10 days in 20%, 3%, 1% oxygen. For culture 1x105cell/ml were seeded in colony forming assay and 2x106cell/ml were seeded in L-glutamin mediain chamber slide. We counted cell colonies under different hypoxic condiontins by Olympus IX71 fluorescence microscope. After cell culture in chamber slide, we stained cells by anti-CD90 and anti-CD105 then counted positive cells by Olympus IX71 fluorescence microscope.
RESULTS: Compared to normoxic cells and hypoxic cells well morphologically differentiated and counted by Olympus IX71 microscope. More colonies were observed at 3%, 1% oxygen. Statistical significances were identified with granulocytes and macrophage colony (p<0.05) in hypoxic condition. More anti-CD90 and anti-CD105 markers were observed at 3% oxygen condition. Statistical significances were identified in 3% oxygen condition with cell markers(p<0.001).
CONCLUSIONS: Our data suggests low physiological oxygen culture could improve the stemness of macrophage and granulocytes colony and improve the differentiation of mesenchymal cells. Long term culturewith additional cell markers will be necessary to confirm whether low physiological oxygen levels also improve genomic stability