Research Of Effects On Proliferation And Differention Of MC3T3-E1 By The Mice Cerebral Cortex Neurons Conditioned Medium

Objective:To observe the effects on proliferation and differentiation of MC3T3-E1 by mice cerebral cortex conditional medium and explore its molecular mechanism.Materials and Methods:Cerebral cortex neuron were primarily cultered from newborn mice within 24 hours. MC3T3-E1 was cultured by different proportions (1:4,1:2,1:1,1.5:1,2:1,4:1) of conditioned medium collected from 7th day to 10th day and DMEM (high glucose). MTT colorimetric assay was used to determine the ability of growth and proliferation. The group of 1:1 and 1:2 were chosen for culturing MC3T3-E1 and observed the cell cycle by flow cytometry. Cytoskeleton was observed by confocal laser scanning microscope. ALP activity was detected by microplate method.Expressions of COL I 、OPN、 BMP-2 were detected by western blot and was observed the calcium by alizarin red stain.Result:(1) Different proportions (1:4,1:2,1:1,1.5:1,2:1,4:1) of mice cerebral cortex neuron conditional medium had different effects on the proliferation of MC3T3-E1. The group of 1:1 promoted most which was better than control group and had statistic significant P<0.05). The cell cycle of experimental group were more in division stage(S%+G2M%) compare to control group (P<0.05). Confocal laser scanning microscope revealed that the fluorescence intensity of cytoskeleton in experimental group higher than control group (P<0.05).(2) ALP activity, COL I,OPN,BMP-2 and the number of mineral node of experiment group were higher than control group and had statistic significant(P<0.05).Conclusion:The mice cerebral cortex neuron can directly promote the proliferation and differentiation.