摘要
目的:分离培养SD大鼠骨髓基质细胞(bonemarrowstromalcebls,BM-SCs),观察BMSCs部分生物学特性。方法:实验于2004-03/08在珠江医院广东省神经医学研究所实验室进行。采用密度梯度离心法分离BMSCs,MTT法绘制细胞生长曲线、流式细胞仪检测细胞生长周期、鉴定细胞表面标志,免疫荧光染色观察细胞的神经分化功能及端粒酶反转录酶(TERP)表达。PCR-ELISA法检测细胞端粒酶活性。结果:①采用密度梯度离心法可以得到高纯度的BMSCs,表达CD29,CD44等表面标志,而不表达CD31,CD34,CD45。②BMSCs主要由Ⅰ,Ⅱ型两种形态的细胞组成,其中Ⅱ型细胞表达端粒酶活性并可以诱导分化为Nestin阳性细胞。③骨髓基质细胞原代培养时增殖能力较低。结论:BMSCs主要有两种形态和功能不完全相同的细胞类型组成,其中Ⅱ型细胞具有端粒酶活性,并在一定的诱导条件下可以分化为Nestin阳性细胞,可以考虑作为神经移植的种子细胞来源。
AIM: To separate and culture bone marrow stroma cells(BMSCs) of rats and observe the partial biological properties of BMSCs. METHODS:This study was finished in Guangdong Institute of Neurosurgery of Zhujiang Hospital in March 2004.The density gradient centrifugation method was used to separate BMSCs.MTT assay was used to draw cellular growth curve,flow cytometer was used to cells cycle estimated cell cycle and signs of cell surface,immunofluorescence was used to observe neural differentiation function and the expression of telomerase reverse transcriptase and PCR ELISA assay was used to estimate the activity of telomerase in this study respectively. RESULTS:①High purity of BMSCs could obtain with density gradient centrifugation method.BMSCs isolated from adult rat experessed some special markers such as CD29 and CD44,but they did not express CD31,CD34 and CD45.②BMSCs were made up of two different morphological types cells-typeⅠand typeⅡ.The telomerase activities were detected in typeⅡcells which could be induced into the cells with positive nestin expression.③The proliferation activity was slow in primary culture of BMSCs. CONCLUSION:BMSCs are made up of two different morphological and functional types cells-typeⅠand typeⅡ.The typeⅡcells can be induced into nestin positive cells in some condition and show the telomerase avtivity.BMSCs can be considered as a source of seed cells used in neural transplantation.
出处
《中国临床康复》
CSCD
北大核心
2005年第5期52-54,i001,共4页
Chinese Journal of Clinical Rehabilitation
基金
国家自然科学基金(3027049)
广东省重大科技项目犤粤企(2001)367
(2003)209
粤科基办(2004)08犦资助
