利用单克隆抗滋养层抗体（NDOG2）通过流式细胞术分离和鉴定人绒毛膜细胞滋养层亚群。

Isolation and characterisation of a subpopulation of human chorionic cytotrophoblast using a monoclonal anti-trophoblast antibody (NDOG2) in flow cytometry.

作者信息

Caulfield J J, Sargent I L, Ferry B L, Starkey P M, Redman C W

机构信息

Harris Birthright Centre for Pre-eclampsia Research, Nuffield Department of Obstetrics and Gynaecology, John Radcliffe Hospital, Oxford, U.K.

出版信息

J Reprod Immunol. 1992 Jan;21(1):71-85. doi: 10.1016/0165-0378(92)90041-2.

DOI:10.1016/0165-0378(92)90041-2

PMID:1734079

Abstract

Human cytotrophoblast cells, isolated from term amniochorion by enzymic digestion and Percoll gradient centrifugation, were characterised by flow cytometry. A panel of 12 anti-trophoblast monoclonal antibodies was screened for labelling of these cells in flow cytometry and the results compared with immunoperoxidase labelling of cytospin preparations and tissue sections. All 12 antibodies were positive for trophoblast on tissue sections, 11/12 were positive on cytospins but only two (NDOG2 and GB25) gave consistent results in flow cytometry. Two-colour labelling with NDOG2 and W6/32, an antibody to HLA-A, -B, -C, demonstrated that 88% of the NDOG2-positive cells also express Class I major histocompatibility complex (MHC) antigens. The NDOG2-positive cytotrophoblast subpopulation was isolated by flow cytometry in sufficient purity (greater than 95%) and yield (3.1 x 10(6)) for use in functional studies in vitro.

摘要

通过酶消化和Percoll梯度离心从足月羊膜绒毛膜中分离出的人细胞滋养层细胞，用流式细胞术进行了表征。筛选了一组12种抗滋养层单克隆抗体，用于在流式细胞术中标记这些细胞，并将结果与细胞涂片制剂和组织切片的免疫过氧化物酶标记结果进行比较。所有12种抗体在组织切片上对滋养层呈阳性，11/12在细胞涂片上呈阳性，但在流式细胞术中只有两种（NDOG2和GB25）给出一致的结果。用NDOG2和W6/32（一种针对HLA-A、-B、-C的抗体）进行双色标记，结果表明88%的NDOG2阳性细胞也表达I类主要组织相容性复合体（MHC）抗原。通过流式细胞术分离出纯度足够高（大于95%）且产量足够（3.1×10⁶）的NDOG2阳性细胞滋养层亚群，用于体外功能研究。