The term basal plate of the human placenta as a source of functional extravillous trophoblast cells

Abstract

BackgroundExtravillous trophoblast (EVT) cells are of pivotal importance in human embryo implantation and homeostasis of the maternal fetal interface. Invasion of the endometrium by EVT contributes to placental anchorage, spiral artery remodeling, immunological defense, tolerogenic responses, and several collaborative cross talks involved in establishing and maintaining a successful pregnancy. We report here an improved protocol for the isolation of fully differentiated EVT cells from the basal plate of the human term placenta.MethodsThe basal plate was carefully dissected from the villous tissue and the amniochorion membrane prior to enzymatic digestion. Term basal EVT cells were isolated using a 30 and 60% Percoll gradient. A panel of markers and characteristics of the isolated cells were used to confirm the specificity and efficiency of the method so that their potential as an investigative tool for placental research could be ascertained.ResultsIsolated cells were immunoreactive for cytokeratin-7 (CK-7), placental growth factor, placental alkaline phosphatase, human leukocyte antigen G1 (HLA-G1), and α1 and α5 integrins, similarly to the EVT markers from first trimester placental villi. Around 95% of the isolated cells labeled positively for CK-7 and 82% for HLA-G1. No significant change in viability was observed during 48 h of EVT culture as indicated by propidium iodide incorporation and trypan blue test exclusion. Genes for metalloproteinases MMP-2 and MMP9 (positive regulators of trophoblast invasiveness) were expressed up to 48 h of culturing, as also the gelatinolytic activity of the isolated cells. Transforming growth factor (TGF)-beta, which inhibits proliferation, migration, and invasiveness of first-trimester EVT cells, also reduced invasion of isolated term EVT cells in transwell assays, whereas epidermal growth factor was a positive modulator.ConclusionsTerm basal plate may be a viable source of functional EVT cells that is an alternative to villous explant-derived EVT cells and cell lines. Isolated term EVT cells may be particularly useful in investigation of the role of trophoblast cells in pathological gestations, in which the precise regulation and interactive ability of extravillous trophoblast has been impaired.

Tumorigenic Factor CRIPTO-1 Is Immunolocalized in Extravillous Cytotrophoblast in Placenta Creta

“…Interestingly, our previous study demonstrated that the extravillous trophoblast cells retained some capacity for migration and invasion, though it was less than in first trimester placentas [37]. These findings reinforce the conclusion that CRIPTO-1, mostly expressed in potentially invasive cells, is associated with this cellular activity.…”

Methotrexate Induces Apoptosis of Postpartum Placental Cytotrophoblasts

“…Cytotrophoblasts are known to have the potential for transforming to the invasive phenotype [Guilbert et al, 2002;Borbely et al, 2014]. Our research showed that PPTC still exhibited invasiveness, which is consistent with earlier findings [Hohn and Denker, 2002;Borbely et al, 2014;Duzyj et al, 2015].…”

“…Our research showed that PPTC still exhibited invasiveness, which is consistent with earlier findings [Hohn and Denker, 2002;Borbely et al, 2014;Duzyj et al, 2015]. However, of note, EVT also express cytokeratin 7, and it is possible that purified cytotrophoblasts contaminated with EVT exhibit invasiveness once incubated in Matrigel-coated Transwell inserts.…”

Single-cell transcriptomics of the human placenta: inferring the cell communication network of the maternal-fetal interface

“…This cluster of cells is unique in its expression of nonclassical human leukocyte antigen HLA-G and integrin 5 alpha ITGA5-both considered markers of extravillous trophoblast, KRT7 (Telugu et al 2013;Borbely et al 2014) and ADAM12 (Biadasiewicz et al 2014). Further EVT-specific genes and other highly expressed genes are listed in Tables 1 and 2 and discussed in the Supplemental Material.…”