Study Of Real-Time Monitoring The Quantity Of Rat Mesenchymal Stem Cells (rMSCs) Proliferation And Differentiation In Vitro

Objective: To explore the possibility of real-time monitoring the number of Rat Bone Mesenchymal Stem Cells (MSCs) proliferating in vitro and the disparity differentiation capacity between static cells and after continue perfusion cultivation cells .Methods: Rat Mesenchymal Stem Cells were semi-continuous cultured on 24 wells flat plate and continuous perfusion cultured on self-made bioreactor individually. Concentration of glucose and lactic acid were determined on Nova Bioprofile 100+ and the correspond cells number on the culture process were determined through CCK-8 method. Static cultivation cells and after continuing perfusion cultivation cells were induced osteogenic differentiation under the influence of 10-8 M dexamethasone, 10mmol/Lβ- glycerol phosphate ,and 50μg/ ml ascorbate and in the presence of 15% v/v FBS. Two weeks later, determinate the activity of alkaline phosphatase and observe alkaline phosphatase staining and alizarin Bordeaux staining. Adipogenic differentiation were induced in the expanded Static cultivation cells and after continuing perfusion cultivation cells cultures by treatment 0.5mmol/L 1-methyl-3–isobutylx- anthin- e ,1μmol/L dexamethasone, and 1μg/ml insulin. Observe the changes of induced cells. Two weeks later , oil red–o stain show adipogenic differentiation .Results: On the extent, glucose consumption rat and lactic acid production rat were Linear relationship with cell density. It was suggested that cell specific metabolic rates are constant for the range of cell densities studied, on semi-continuous 24 wells flat plate cultivation, qGlu=1.84 mg/ 106 cells-day , qLac=1.74mg/106cells-day and on self-made bioreactor continuous perfusion cultivation, qGlu=1.66 mg/106 cells-day qLac=1.61 mg/106cells-day. there is no obvious differentiation capacity disparity between static cultivation cells and after continue perfusion cells .Conclusion: Real-time monitoring proliferation in vitor MSCs concentration is feasible . there is no obvious differentiation capacity disparity between static cultivation cells and after continue perfusion cells .