Project description:The goal of this study was to identify the gene expression signatures of two closely related types of trophoblast in human placentas: villous cytotrophoblasts (vCTB) and proximal column extravillous trophoblasts (pcEVT). The two populations were isolated from first trimester placentas and identified using the specific surface markers, EGFR (vCTB) and HLA-G (pcEVT).
Project description:Comparison of genes associated with the EMT between cytotrophoblast cells (CTB) and extravillous trophoblast cells (EVT) from normal third trimester placenta and abnormally invasive placenta (AIP)
Project description:The goal of the present study was to characterize HLA-G-positive cells derived from human embryonic stem cells treated with BMP4 for five days and compare them to HLA-G positive cells derived from human first trimester placentas
Project description:Human trophoblast stem cells (hTSCs) have emerged as a powerful tool for modeling the placental cytotrophoblast (CTB) in vitro. hTSCs were originally derived from CTBs of the first trimester or blastocyst-stage embryos in trophoblast stem cell medium (TSCM) that contains epidermal growth factor (EGF), the glycogen synthase kinase-beta (GSK3b) inhibitor CHIR99021, the transforming growth factor-beta (TGFb) inhibitors A83-01 and SB431542, valproic acid (VPA), and the Rho-associated protein kinase (ROCK) inhibitor Y27632. Here we show that hTSCs can be derived from CTBs isolated from the term placenta, using TSCM supplemented with a low concentration of mitochondrial pyruvate uptake inhibitor UK5099 and lipid-rich albumin (TUA medium). Notably, hTSCs could not be derived from term CTBs using TSCM alone, or in the absence of either UK5099 or lipid-rich albumin. Strikingly, hTSCs cultured in TUA medium for a few passages could be transitioned into TSCM and cultured thereafter in TSCM. hTSCs from term CTBs cultured in TUA medium as well as those transitioned into and cultured in TSCM thereafter could be differentiated to the extravillous trophoblast and syncytiotrophoblast lineages, and exhibited high transcriptome similarity with hTSCs derived from first trimester CTBs. We anticipate that these results will enable facile derivation of hTSCs from normal and pathological placentas at birth with diverse genetic backgrounds, and facilitate in vitro mechanistic studies in trophoblast biology.
Project description:During pregnancy, trophoblast cells in the placenta are the only fetal cells in direct contact with the maternal blood and decidua. They have many functions, including transport of nutrients and oxygen, remodelling the uterine arteries, and communicating with maternal cells. Despite their importance in development and in the success of pregnancy, little is known about human trophoblast progenitors and their differentiation. We identify a proliferative trophoblast niche at the base of cytotrophoblast cell columns in first trimester placentas that is characterised by integrin α2 (ITGA2) expression. Pulse-chase experiments with 5-Iodo-2′-deoxyuridine (IdU) imply that these cells can contribute to both villous and extravillous lineages. Importantly, these cells can be isolated by ITGA2 using flow cytometry and express genes from both VCT and EVT. Microarray shows that they display a unique gene signature including NOTCH signalling and mesenchymal characteristics. ITGA2 allows for the first time the study of a pure population of trophoblast progenitor cells.
Project description:Comparison of genes associated with the EMT between undifferentiated cytotrophoblast cells (CTB) and differentiated extravillous trophoblast cells (EVT) from third trimester human placenta. Cells isolated from control (placenta previa) and cases (preeclampsia). Cells isolated by immunomagnetic separation using anti-integrin beta4 antibody to purify CTB and anti-HLA-G antibody to purify EVT.
Project description:To better understand how DNA methylation influences placentation, DNA from first trimester primary trophoblast populations (side-population trophoblasts, cytotrophoblasts and extravillous trophoblasts) isolated using FACS underwent reduced representation bisulfite sequencing and were compared to publicly available data of blastocyst-derived and somatic cell populations.
Project description:In this work, we have isolated a Hoescht side-population of trophoblasts from first trimester human placentae that cluster separately from more differentiated trophoblast populations, and have a transcriptomic profile indicative of a stem cell population. Hoescht side-population cells were compared in quintuplicate with extravillous trophoblasts and cytotrophoblasts extracted from the same placentae, giving a total of 15 samples.
Project description:The goal of the present study was to characterize HLA-G-positive cells derived from human embryonic stem cells treated with BMP4 for five days and compare them to HLA-G positive cells derived from human first trimester placentas Human embryonic stem cells were cultured with BMP4 in the absence of FGF2 for 5 days. They were then dispersed and separated into HLA-G+ and HLA-G-, TACSTD2 positive cells by using immunomagnetic bead separation. First trimester human placental samples from 8-12 weeks gestation were enzymatically separated into single cells, and then also separated into HLA-G+ and HLA-G-/TACSTD2+ cell populations
