糖浓度对三维培养体系下小鼠胚胎干细胞成骨分化的影响

Effect of Glucose Level on Osteogenic Differentiation of Mouse Embryonic Stem Cells in 3D Culture System

目的:探讨明胶海绵作为小鼠胚胎干细胞(ESCs)成骨诱导支架材料的可行性,并考察葡萄糖浓度对三维培养体系下小鼠ESCs骨向分化的影响。方法:体外培养小鼠ESCs,将消化所得的单细胞接种到可降解明胶海绵上,分别于高糖骨诱导培养液和低糖骨诱导培养液中进行培养,同时设立自然分化对照组。结果:诱导后第7、14天茜素红染色结果显示,高糖组矿化结节数少于低糖组。结论:明胶海绵可以作为小鼠胚胎干细胞体外成骨诱导的培养支架;在骨诱导过程中高葡萄糖浓度抑制了小鼠胚胎干细胞的骨向分化能力。

Objective： To explore the possibility of gelatin sponge as a scaffold of mouse embryonic stem ceils （ESCs） in vitro cultivation and to investigate the effect of glucose level on osteogenic differentiation of mouse ESCs. Methods： Mouse ESCs were cultured in vitro, and the dispersed cells were implanted into absorbable gelatin sponge for osteogenic induction. Three groups were set in the study： high-glucose-level group, low-glucose-level group and spontaneous differentiation group. After 7 and 14 days of osteogenic differentiation, the matrix minerali- zation was detected by alizarin red S staining. Results： After 7 and 14 days of osteogenic induction, low-glueose- level group had a higher percentage of alizarin red S positive cells, compared with high-glucose-level group and spontaneous differentiation group（P〈0. 05）. Conclusion： Gelatin sponge can be used as a scaffold for culturing mouse ESCs in vitro study. High glucose level inhibits osteogenetic differentiation of mouse ESCs in the 3D culture system.