用于高场MRS研究的人脐带间充质干细胞代谢物提取方法选择的优化  

Extraction methods optimization in high-field MRS study of the human umbilical cord mesenchymal stem cells

作　　者：代海洋[1] 洪璧楷[1] 肖叶玉[1] 杨玉萍[2] 马廉[2] 陈耀文[3] 吴仁华[1] 

机构地区：[1]汕头大学医学院第二附属医院影像科,广东汕头515041 [2]汕头大学医学院第二附属医院儿科,广东汕头515041 [3]汕头大学中心实验室,广东汕头515041

出　　处：《磁共振成像》2011年第6期430-434,共5页Chinese Journal of Magnetic Resonance Imaging

基　　金：国家自然科学基金重点项目(编号:30930027);国家自然科学基金面上项目(编号:60971075);汕头大学医学院基础与临床科研基金项目(2010年)支持

摘　　要：目的从代谢物提取效率及9.4TMRS对不同提取物检测敏感性差异的角度分析比较离体频谱研究中常用的代谢物提取方法,对提取方法的选择进行一定的优化。方法培养获取人脐带间充质干细胞,利用高氯酸(perchloric acid,PCA)提取细胞水溶性代谢物,利用甲醇/氯仿(methanol-chlo roform,M/C)提取细胞脂溶性代谢物,并使用甲醇/氯仿/水一次性提取方法(dual phase extraction,DPE)同时获取两种组分,对不同方法所得提取物干重进行配对样本t检验。利用9.4T磁共振获取不同干重的水溶性及脂溶性提取物频谱,寻找能获得良好谱线质量的提取物重量。结果相比于DPE法,PCA对水溶性代谢物具有更高的提取效率,M/C对脂溶性代谢物具有更高的提取效率。高场MRS能更敏感地检测出低浓度的脂溶性物质,不足1mg的脂溶性提取物就能得到良好的谱线质量,而约6mg的水溶性提取物才能得到较好的谱线。结论可以利用PCA提取水溶性代谢物联合M/C提取脂溶性代谢物,在获得更多代谢信息的同时达到对细胞的充分利用。Objective： To explore the suitable cell extraction methods that most fit in vitro MRS study by analyzing the extraction efficiencies and detecting sensitivity, and to make some optimizations to the choice of different extraction procedures. Materials and Methods： Human umbilical cord mesenchymal stem cells were cultured and collected. The water-soluble metabolites were extracted by perchloric acid （PCA） and the lipid-soluble metabolites were extracted by methanol-chloroform （M/C）, and we employed a dual phase extraction （DPE） procedure by methanol-chloroform-water to get both fractions from the same cell batch. The net weights from different extraction methods were compared and analyzed using Student paired t-test. Results： Compared with the net yields from DPE, the PCA extraction procedure has a higher efficiency in extracting water-soluble metabolites while M/C is efficient in extracting lipid-soluble metabolites. MRS is highly sensitive in detecting extremely small quantity of lipid-soluble metabolites. Conclusion： We can employ PCA combined with M/C to obtain both water- and lipid-soluble metabolites for the efficient use of cells in MRS analysis.

关 键 词：间充质干细胞 磁共振频谱 代谢物提取 优化 

分 类 号：R445.2[医药卫生—影像医学与核医学]