摘要
目的探讨三磷酸胞苷二钠(CTP)诱导骨髓间充质干细胞分化为神经元样细胞的作用。方法取成年SD大鼠,用贴壁培养法分离纯化骨髓间充质干细胞(BMSCs),传代扩增,连续观察细胞形态及生长特性。用不同浓度(0.05、0.10、0.20、0.40和0.80mg/mL)CTP对BMSCs进行诱导;诱导后用免疫组织化学检测细胞神经微丝蛋白(NF)、胶质原性纤维酸性蛋白(GFAP)和神经元烯醇化酶(NSE)的表达。计算阳性细胞比例,进行统计学分析,选出合适的诱导剂量。结果随CTP浓度增高,经免疫组织化学检测NF、GFAP和NSE阳性细胞比例也呈现逐步增高趋势,到一定浓度后又有所下降,统计学分析,CTP浓度为0.2mg/mL时,能够诱导NF、GFAP和NSE阳性细胞比例为(32.73±0.63)%、(62.77±2.45)%和(30.41±0.65)%,显著高于对照组(P<0.01)。结论 CTP是一种很好的体外诱导剂,可以诱导MSCs向神经细胞分化,以0.2mg/mLCTP作诱导剂,能够获得较明显的诱导MSCs向神经细胞和神经胶质细胞分化的结果。
[Objective] To investigate whether BMSCs could be induced to differentiate into neuron-like cells by Cytidine Triphosphate （CTP）. [Methods] BMSCs were obtained from femurs of Sprague-Dawley adult rat under anesthesia. BMSCs were purified from the bone marrow by density gradient centrifugation and by adhering to the culture flasks. Cell morphologic change and growth property were obtained continuously. BMSCs was induced by 5 different concentration of CTP （0.05 mg/mL, 0.1 mg/mL, 0.2 mg/mL, 0.4 mg/mL, 0.8 mg/ mL）, the ratio which BMSCs differentiated into neural cells was examined through neurofilament protein （NF）, gliad fibrillary acidicprotein （GFAP） and neuronspecific enolase （NSE） by immunohistoehemistry assays. [Resuits] Compared with control groups, positive cell in NF, GFAP and NSE increased when CTP concentration from 0.05 mg/mL to 0.8 mg/mL by immunohistochemistry assays and statistics. The positive expression of NF, GFAP and NSE were （32.73±0.63）%, （62.77±2.45）%, （30.41±0.65）% respectively when CTP （0.2 mg/mL） in duced. [Conclusion] CTP can induce BMSCs into neuron-like cells in vitro. We conclude that CTP （0.2 rag/ mL） which was the optimal concentration.
出处
《中国现代医学杂志》
CAS
CSCD
北大核心
2011年第17期1970-1973,1976,共5页
China Journal of Modern Medicine
基金
河北省保定市科技局科学技术研究与发展计划项目(No:06F04-11)
河北省卫生厅科研基金项目(No:20090175)
河北省中医药管理局科研计划项目(No:2009153)
