CD1332/AC141——一种滋养层细胞新的细胞内标志  

CD133-2/AC141: a novel intracellular marker for human trophoblast

作　　者：杜美蓉[1] 李大金[1] 王明雁[1] 朱影[1] 袁敏敏[1] 孟毅[1] 

机构地区：[1]复旦大学妇产科研究所,上海200011

出　　处：《生殖医学杂志》2004年第6期360-364,共5页Journal of Reproductive Medicine

基　　金：上海市基础研究重点项目基金资助 (0 3JC14 0 16);复旦大学"985工程"项目 (985B3 6);计划生育药具国家重点实验室课题资助 (B2 0 2 0 1)

摘　　要：目的　了解CD133在滋养细胞的表达情况 ,为鉴定滋养细胞株及原代分离的滋养细胞提供新的细胞内标志。　方法　通过胰蛋白酶 /DNaseⅠ分步消化 ,Percoll密度梯度离心分离纯化滋养细胞 ,经抗角蛋白 (CK 7)、波形蛋白 (vimentin)单克隆抗体行免疫细胞化学和流式细胞仪 (FACS)鉴定 ,纯度达 95 %以上。采用三种不同的CD133荧光标记单克隆抗体在70 %的甲醇固定前后对原代分离的滋养细胞和绒毛膜上皮癌细胞株 (JAR)行FACS分析。　结果　 70 %甲醇固定前 ,三种CD133单克隆抗体均不能与滋养细胞反应 ;固定后 ,抗CD133 1(AC133 1 APC)、抗CD133 2 (2 93C3 PE)仍不能与滋养细胞反应 ,而抗CD133 2 (AC14 1 PE)与滋养细胞胞内抗原结合。而且AC14 1阳性的细胞也是CK 7阳性的细胞。　结论　CD133 2 /AC14 1可作为滋养细胞纯度鉴定的一种有用标志 ,但应用时应选择合适的抗体。Objective: To provide a new intracellular marker to characterize human trophoblastic cell.Methods: Human first-trimester trophoblastic cells were isolated by trypsin/DNase I digestion and discontinuous Percoll density gradient centrifugation. Identified by immunocytochemistry and FACS with monoclonal antibody (mAb) to cytokeratin, the trophoblastic cells were more than 95% pure. The CD133 expression in the trophoblast and JAR cells was analyzed by FACS after methanol fixation. Results: None of the three monoclonal antibodies (CD133-1, CD133-2, CD133-2/AC141) to CD133 can react with primary trophoblast and JAR cells before methanol fixation. After fixed with methanol, only CD133-2 mAb (clone number AC141) can react with human trophoblastic cells. The AC141-positive cells are also cytokeratin7- positive. Conclusion: The CD133-2/AC141 can be used as a useful marker to identify the purity of primary trophoblastic cells.

关 键 词：CD133-2 人滋养细胞 

分 类 号：Q813.11[生物学—生物工程]