摘要
目的探讨三七总皂甙(tPNS)诱导骨髓间充质干细胞(MSCs)分化为神经元样细胞过程中细胞内钙离子浓度([Ca2+]i)的变化。方法LDMEM冲洗SD大鼠股骨骨髓腔,培养大鼠MSCs。选用第5代MSCs进行诱导分化,用含10μg/L碱性成纤维细胞生长因子(bFGF)的LDMEM培养液预诱导24h,加入含10μmol/LFluo3/AM的LDMEM培养液负载30min,最后更换成含三七总皂甙0.25g/L的无血清培养液,同时在波长为488nm激光下扫描观测细胞形态和荧光强度的变化。结果更换三七总皂甙诱导液后,细胞内荧光强度逐渐增加,到100s达高峰值,其后逐渐减弱,但20min时细胞荧光强度仍高于诱导前,此时可见MSCs开始向神经元样细胞分化。结论三七总皂甙在体外诱导MSCs分化为神经元样细胞过程中胞内游离钙离子[Ca2+]i浓度升高。
Objective To investigate the changes in the concentration of calcium ion _i in mesenchymal stem cells during their differentiating into neuron-like cells induced by total panax notoginseng in vitro. Methods Rat MSCs were separated from femur marrow by flushing out with low glucose DMEM and then cultured. After 5 passages in culture,MSCs were pre-induced by 10 g/L bFGF for 24h and then, with the addition of L-DMEM containing 10 mol fluro-3/AM, for another 30min. Last, the medium was replaced with an induction medium containing 0.25g/L tPNS. Meanwhile, we observed the changes of the fluorescence intensity and the shape of MSCs with laser beams of 488 nm wavelength. Results The fluorescence intensity in MSCs increased gradually after the medium was replaced with the induction medium containing tPNS. At 100s it attained its peak value and then decreased gradually, but at min still higher than that before the induction. Meanwhile, we observed that MSCs differentiated into neuron-like cells. Conclusion Calcium ion_i increased in MSCs during differentiating into neuron-like cells.
出处
《中国组织化学与细胞化学杂志》
CAS
CSCD
2005年第3期241-244,共4页
Chinese Journal of Histochemistry and Cytochemistry
基金
国家重点基础研究课题(973)资助项目(G19990543012)
广东省科委资助项目(2004B33801003)
关键词
骨髓间充质干细胞
神经元
三七总皂甙
细胞内游离钙离子
Mesenchymal stem cell
Neuron
Total panax notoginseng saponins
Intracellular free calcium ion
