用于检测细胞粘附的^3H—TdR掺入实验  

Application of 3 H-TdR incorporation assay in the detection of cell-cell adhesion

作　　者：孙凯[1] 金伯泉[2] 冯琦[3] 朱勇[2] 杨琨[2] 刘雪松[2] 董帮权[2] 

机构地区：[1]第四军医大学西京医院肝胆外科,陕西西安710032 [2]第四军医大学免疫学教研室 [3]第四军医大学西京医院血液内科

出　　处：《西北国防医学杂志》2002年第4期241-243,共3页Medical Journal of National Defending Forces in Northwest China

基　　金：国家自然科学基金资助项目 ( 3970 0 0 6 5 )

摘　　要：目的 :细胞与细胞之间及细胞与基质之间的粘附作用在细胞间的信息传递中起极其重要的作用 ,检测细胞间粘附作用是细胞生物学研究中不可缺少的方法之一。方法 :我们建立了以3 H -TdR掺入实验为基础的细胞粘附检测方法。①首先将待检测细胞之一于 96孔培养板培养使之贴壁 ,进行细胞铺底 ;②将另一待检细胞标记3 H -TdR ;③将 2种细胞在培养板中共同孵育 4h ,使之相互作用 ;④轻柔洗去未粘附细胞 ,将剩余细胞以多头细胞收集仪收集 ,β计数仪测cpm值 ;⑤可通过单抗等进一步证实细胞间粘附的特异性。结果 :将3 H -TdR掺入实验用于细胞粘附的检测具有较高的敏感性 ,它能准确地检测细胞间的粘附现象 ,并反映其相对强弱。结论 :3 H -TdR掺入实验由于采用同位素标记方法 ,敏感性较好。通过阻断实验亦能保证实验的特异性 。Objective:The adhesion of cells to cells and cells to stroma play very important role in the cell-cell information exchanges.Methods:The cell adhesion assay based on the 3 H-TdR incorporation assay was introduced. Firstly, coated cells were cultured to confluence in 96 well plate. After incubated with 3 H-TdR for 6h, the isotope labeled cells were added into plate wells and incubated for another 4 h. Then the un-adhered cells were removed by gently washing. The cpm was counted after cell harvest.Results:The cell adhesion assay based on the 3 H-TdR incorporation assay was very sensitive in the detection of cell adhesion. It could reflect the relative of cell to cell adhesion.Conclusion:The cell adhesion assay based on the 3 H-TdR incorporation assay is an useful method in the detection of cell to cell adhesion.

关 键 词：实验室诊断 检测 细胞粘附 同位素 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]