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The human placenta provides the site of exchange between the maternal and fetal bloodstreams, acts as an endocrine organ, and has immunological functions. The majority of pregnancy disorders including preeclampsia and fetal growth restriction have their roots in pathological placentation. Yet, the underlying molecular causes of these complications remain largely unknown, not least due to the lack of reliable in vitro models. Recent establishment of 2D human trophoblast stem cells and 3D trophoblast organoids has been a major advancement that opened new avenues for trophoblast research. Here we provide a protocol detailing isolation of cytotrophoblast from the first trimester human placenta, establishment of trophoblast organoids, their culture and differentiation conditions. Overall, we describe an in vitro system that offers an excellent model to study the molecular basis of placental development and disease.
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http://dx.doi.org/10.1007/978-1-0716-3495-0_17 | DOI Listing |
NAR Mol Med
July 2025
Department of Biochemistry and Molecular Biology, Cumming School of Medicine, University of Calgary, 3330 Hospital Drive NW, Calgary, Alberta T2N 4N1, Canada.
Advanced maternal age increases the risk of pregnancy complications due, in part, to changes in the uterine environment. Here, we show that uterine aging in mice is associated with a progressive increase in transcriptional variation, accompanied by a notable accumulation of activating histone marks at multiple genomic loci. Importantly, the transcriptional signatures of uterine aging differ substantially from senescence markers associated with organismal aging.
View Article and Find Full Text PDFDevelopment
September 2025
Department of Developmental Biology, Washington University in St. Louis, School of Medicine, St. Louis, MO 63110, USA.
CCR7 chemokine G protein-coupled receptor is expressed in extraembryonic tissues of the early human embryo, including trophectoderm and its derivatives cytotrophoblast (CTB), extravillous trophoblast (EVT), and syncytiotrophoblast (STB). However, its function in placentation remains understudied. Here, we generated human embryonic stem cells harboring CCR7 deletions and differentiated them into human trophoblast stem cells (hTSC), their EVT and STB derivatives, and trophoblast organoids.
View Article and Find Full Text PDFPLoS Biol
August 2025
Department of Integrative Immunobiology, Duke University School of Medicine, Durham, North Carolina, United States of America.
The placenta is a dynamic, embryo-derived organ essential for fetal growth and development. While all eutherian mammals have placentas composed of fetal-derived trophoblasts that mediate maternal-fetal exchange, their anatomical and histological structures vary across species due to evolutionary divergence. Despite the cellular heterogeneity of porcine trophoblasts in vivo, understanding the mechanisms driving porcine placental development has been limited by the lack of in vitro models replicating this heterogeneity.
View Article and Find Full Text PDFNat Cell Biol
August 2025
Center for Life Sciences, School of Life Sciences, Yunnan University, Kunming, China.
The first embryonic cell differentiation in mice segregates the trophectoderm and the inner cell mass. Successful derivation of mouse trophoblast stem cells (TSCs) and trophectoderm stem cells (TESCs) has greatly facilitated the understanding of trophoblast differentiation. However, our understanding of early trophectoderm differentiation remains incomplete.
View Article and Find Full Text PDFBackground: The placenta is a temporary organ that develops throughout pregnancy, connecting a developing fetus to the maternal uterine wall. The placenta's structure is species specific and complex, resulting in recent advancements with in vitro models to help study this dynamic organ. The main cell type composing the placenta, trophoblast cells, serve several roles and have been incorporated within biomaterials and devices to recapitulate the placental microenvironment.
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