Isolation and characterization of human trophoblast side-population (SP) cells in primary villous cytotrophoblasts and HTR-8/SVneo cell line.
Isolation and characterization of human trophoblast side-population (SP) cells in primary villous cytotrophoblasts and HTR-8/SVneo cell line.
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DOI：
10.1371/journal.pone.0021990
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发表时间：
2011
期刊：
影响因子：
3.7
通讯作者：
Wake N
中科院分区：
文献类型：
--
作者：
Takao T;Asanoma K;Kato K;Fukushima K;Tsunematsu R;Hirakawa T;Matsumura S;Seki H;Takeda S;Wake N
最近，许多研究已经确定，包括干细胞和祖细胞在内的未成熟细胞群可以在“侧群”（SP）细胞中找到。虽然从一些成人组织中分离的SP细胞已在其他地方报道，但人滋养层SP的分离和表征仍有待报道。在这项研究中，HTR-8/SVneo细胞和人原代绒毛细胞滋养层细胞（vCTB）染色Hoechst 33342和SP和非SP（NSP）部分使用细胞分选仪分离。在HTR-8/SVneo细胞和vCTB中鉴定出SP细胞的小群体。SP细胞表达几种vCTB特异性标志物，但不表达合体滋养层（STB）或绒毛外细胞滋养层（EVT）特异性分化标志物。SP细胞在小鼠胚胎成纤维细胞（MEF）饲养细胞上或在添加肝素/FGF 2的MEF条件培养基中形成集落并增殖，并且它们也显示出长期的再增殖特性。SP细胞可分化为STB和EVT细胞系，并表达多种分化标志物。基因芯片分析显示，IL 7 R和IL 1 R2只在SP细胞中表达，而在NSP细胞中不表达。被分选为对IL 7 R和IL 1 R2均呈阳性的vCTB细胞不能表达滋养层分化标志物，并且在基础培养基中自发地分化成STB和EVT。SP细胞显示的这些特征表明IL 7 R和IL 1 R2可用作检测SP细胞的标志物，并且vCTB祖细胞和滋养层干细胞参与SP细胞群体。
Recently, numerous studies have identified that immature cell populations including stem cells and progenitor cells can be found among “side-population” (SP) cells. Although SP cells isolated from some adult tissues have been reported elsewhere, isolation and characterization of human trophoblast SP remained to be reported. In this study, HTR-8/SVneo cells and human primary villous cytotrophoblasts (vCTBs) were stained with Hoechst 33342 and SP and non-SP (NSP) fractions were isolated using a cell sorter. A small population of SP cells was identified in HTR-8/SVneo cells and in vCTBs. SP cells expressed several vCTB-specific markers and failed to express syncytiotrophoblast (STB) or extravillous cytotrophopblast (EVT)-specific differentiation markers. SP cells formed colonies and proliferated on mouse embryonic fibroblast (MEF) feeder cells or in MEF conditioned medium supplemented with heparin/FGF2, and they also showed long-term repopulating property. SP cells could differentiate into both STB and EVT cell lineages and expressed several differentiation markers. Microarray analysis revealed that IL7R and IL1R2 were exclusively expressed in SP cells and not in NSP cells. vCTB cells sorted as positive for both IL7R and IL1R2 failed to express trophoblast differentiation markers and spontaneously differentiated into both STB and EVT in basal medium. These features shown by the SP cells suggested that IL7R and IL1R2 are available as markers to detect the SP cells and that vCTB progenitor cells and trophoblast stem cells were involved in the SP cell population.
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影响因子：
3.8
作者：
IRVING, JA;LYSIAK, JJ;LALA, PK
通讯作者：
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通讯作者：
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作者：
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作者：
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