Placenta · 2025
Establishment of human trophoblast stem cells from term smooth chorion.
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Hoshiyama Takako, Muto Masanaga, Matsumoto Shoma, Okamura Eiichi, Jargalsaikhan Bat-Erdene, Murakami Takashi, Tsuji Shunichiro, Ema Masatsugu
Аннотация
INTRODUCTION: Human trophoblast stem (TS) cells derived from first-trimester placental villi (TSCT) and blastocyst (TSblast) are valuable for studying placental development and pathobiology. However, most placenta-mediated pregnancy complications are diagnosed in the third-trimester, and there are limited reports on TS cells from the third-trimester placental tissues. In this study, we report the successful derivation of TS cells from the term smooth chorion.
METHODS: Smooth chorion isolation was carried out from term placenta, followed by the preparation of single cells through enzymatic dissociation. ITGA6-positive cells were isolated from smooth chorion and cultured in TS medium. The chorion-derived TS (Ch-TS) cells were characterized through flow cytometry, immunocytochemical analysis, DNA methylation analyses via bisulfite sequencing, miRNA analysis using quantitative PCR, and RNA sequencing. To assess their differentiation potential, Ch-TS cells were induced to differentiate into extravillous trophoblast (EVT) cells and syncytiotrophoblast (ST) cells.
RESULTS: Human TS cells were successfully derived from the term smooth chorion. Immunohistochemistry confirmed trophoblast marker expression in Ch-TS cells. Bisulfite sequencing revealed that the ELF5 promoter region was hypomethylated in Ch-TS cells, consistent with trophoblastic DNA methylation status. Ch-TS cells expressed miRNAs from the chromosome 19 miRNA cluster (C19MC). Flow cytometryanalysis revealed that the expression patterns of HLA class I molecules were comparable between Ch-TS and TSCT/blast. Ch-TS cells successfully differentiated into EVT and ST cells. RNA sequencing showed transcriptomic similarities between Ch-TS cells and TSCT/blast cells.
DISCUSSION: Ch-TS cells may serve as a valuable in vitro model for studying trophoblast biology and placenta-mediated pregnancy complications, similar to TSCT/blast cells.
METHODS: Smooth chorion isolation was carried out from term placenta, followed by the preparation of single cells through enzymatic dissociation. ITGA6-positive cells were isolated from smooth chorion and cultured in TS medium. The chorion-derived TS (Ch-TS) cells were characterized through flow cytometry, immunocytochemical analysis, DNA methylation analyses via bisulfite sequencing, miRNA analysis using quantitative PCR, and RNA sequencing. To assess their differentiation potential, Ch-TS cells were induced to differentiate into extravillous trophoblast (EVT) cells and syncytiotrophoblast (ST) cells.
RESULTS: Human TS cells were successfully derived from the term smooth chorion. Immunohistochemistry confirmed trophoblast marker expression in Ch-TS cells. Bisulfite sequencing revealed that the ELF5 promoter region was hypomethylated in Ch-TS cells, consistent with trophoblastic DNA methylation status. Ch-TS cells expressed miRNAs from the chromosome 19 miRNA cluster (C19MC). Flow cytometryanalysis revealed that the expression patterns of HLA class I molecules were comparable between Ch-TS and TSCT/blast. Ch-TS cells successfully differentiated into EVT and ST cells. RNA sequencing showed transcriptomic similarities between Ch-TS cells and TSCT/blast cells.
DISCUSSION: Ch-TS cells may serve as a valuable in vitro model for studying trophoblast biology and placenta-mediated pregnancy complications, similar to TSCT/blast cells.