Antibody reaction patterns in first trimester placenta: Implications for trophoblast isolation and purity screening
Antibody reaction patterns in first trimester placenta: Implications for trophoblast isolation and purity screening
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DOI：
10.1053/plac.2000.0559
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发表时间：
2000-09-01
期刊：
影响因子：
3.8
通讯作者：
Desoye, G
中科院分区：
文献类型：
--
作者：
Blaschitz, A;Weiss, U;Desoye, G
免疫组织化学和细胞化学研究的目的是筛选特异的抗体，以建立一种有效的早孕滋养层细胞纯化方案，并用于随后分离的滋养层细胞的纯度筛选。对胎盘组织冰冻切片中各种细胞角蛋白细丝、糖蛋白CD9、成纤维细胞特异性抗原(FSA)、普通白细胞抗原CD45RB、巨噬细胞抗原CD163、CD68和CD14的抗体反应性进行了研究。在所检测的细胞角蛋白中，细胞角蛋白7是唯一的角蛋白细丝类型，在胎盘间充质细胞中不表达，但在所有滋养细胞亚群中都有表达。由于除间充质细胞外，抗CD9抗体也能强烈标记绒毛外细胞滋养层细胞，而抗FSA抗体仅与间充质细胞发生反应，因此抗FSA抗体适合作为间充质细胞的耗竭抗体。在巨噬细胞标志物中，抗CD163对Hofbauer细胞的特异性最强。CD45RB在母体和胎儿白细胞及Hofbauer细胞上均有表达。从密度梯度中收集的分离的妊娠早期胎盘细胞制剂中含有高达45%的非滋养细胞。用CK7、FSA、Vimentin、CD45RB和CD163的抗体进行免疫细胞化学检测表明，CD45RB和FSA的免疫去除使滋养层细胞的纯度提高到98%以上。根据这项研究，早期胎盘滋养层细胞应该用针对异构体7的细胞角蛋白抗体进行鉴定。仅通过密度梯度纯化分离的滋养层细胞不能获得足够程度的纯度。(C)2000 HarCourt出版有限公司。
The aim of this immunohistochemical and cytochemical study was to select specific antibodies to establish an efficient purification protocol for first trimester trophoblast and for subsequent purity screening of isolated trophoblast cells. The reactivity of antibodies to various cytokeratin filaments, glycoprotein CD9, fibroblast specific antigen (FSA), common leukocyte antigen CD45RB and macrophage antigens CD163, CD68 and CD14 were studied on cryosections of placental tissue. Among the cytokeratins tested, cytokeratin 7 was the only keratin filament type, which was not expressed in placental mesenchymal cells, but in all trophoblast subpopulations. Since anti-CD9, in addition to mesenchymal cells, also strongly labels extravillous cytotrophoblast cells, whereas the antibody to FSA only reacts with mesenchymal cells, anti-FSA is suitable as a depletion antibody for mesenchymal cells. Among the macrophage markers anti-CD163 was the most specific for Hofbauer cells. CD45RB was expressed on maternal and fetal leukocytes as well as on Hofbauer cells. Isolated first trimester placental cell preparations that have been collected from a density gradient contained up to 45 per cent non-trophoblast cells. Immunocytochemistry using antibodies to CK7, FSA, vimentin, CD45RB and CD163 demonstrated that subsequent immunodepletion with antibodies to CD45RB and FSA increased the purity of the trophoblast preparation to greater than 98 per cent.According to this study trophoblasts from first trimester placentae should be identified by cytokeratin antibodies specific for the isoform 7. Purification of isolated trophoblasts by density gradient alone does not result in a sufficient degree of purity. (C) 2000 Harcourt Publishers Ltd.