An in vitro model of human placental trophoblast deportation/shedding

Abstract

Deportation of trophoblast shed from the placenta into the maternal circulation was first described over 100 years ago. Despite this, little is known about the quantity or nature of the shed and deported trophoblasts. Neither do we have a clear understanding of the fate of deported trophoblasts nor do we have a clear understanding of their effects on the maternal physiology. This deficiency is largely due to the inaccessibility of deported trophoblasts in vivo. This study aimed to produce a model that would allow us to study deported trophoblasts. We devised a system for culturing placental explants of 12-week gestation in cell culture inserts with a stainless steel mesh bottom that allowed the ready harvesting of shed/deported trophoblasts. Immunohistochemical and morphologic investigations demonstrated that these in vitro shed/deported trophoblasts are similar to those found in vivo and that apoptotic, necrotic and viable trophoblasts were shed from the explants. Inhibiting caspases induced a change from predominantly apoptotic to predominantly necrotic trophoblast shedding. We have devised an in vitro model that allows the collection of shed/deported trophoblasts which will significantly enhance our ability to study these cells. Our preliminary investigations confirm that apoptosis plays an important role in trophoblast shedding/deportation.

Differential Proteomic Analysis of Syncytiotrophoblast Extracellular Vesicles from Early-Onset Severe Preeclampsia, using 8-Plex iTRAQ Labeling Coupled with 2D Nano LC-MS/MS

“…Both methods are widely used [20,23,41,42]. It has previously shown that the syncytiotrophoblast undergoes significant artefactual degradation during the first 24 h of villous explant culture [43]. By 72h of culture, the underlying cytotrophoblast have fused to form a new syncytium that has formed under the syncytium in vitro [43,44].…”

“…It has previously shown that the syncytiotrophoblast undergoes significant artefactual degradation during the first 24 h of villous explant culture [43]. By 72h of culture, the underlying cytotrophoblast have fused to form a new syncytium that has formed under the syncytium in vitro [43,44]. This may explain that Redman et al who only cultured 24h in their explant culture model or used a short term perfusion model to isolate STBM, where the syncytium would have been more representative of that present in vivo.…”

“…This may explain that Redman et al who only cultured 24h in their explant culture model or used a short term perfusion model to isolate STBM, where the syncytium would have been more representative of that present in vivo. But Abumaree et al also considered that the cultured time extend to 48 or 72 h in the explant culture model might be acceptable [43]. We adopted the eSTBM method combined with differential centrifugation, and the STBM preparations were verified by electron microscopy (EM) and western blot analysis.…”

Detection of Fetal Sex, Aneuploidy and a Microdeletion from Single Placental Syncytial Nuclear Aggregates

“…[29] . A , B Optical sections obtained by confocal microscopy demonstrating examples of SNAs shed from non-stained placental explants.…”

“…Placental explants were cultured overnight following an established in vitro model [29] . Trophoblast debris was collected and transferred into 100 × 20 mm BD Optilux TM petri dishes (Becton Dickinson, Auckland, New Zealand) and visualised using either an ELWD 0.3 (Nikon, Auckland, New Zealand) or an Eclipse Ti 50 microscope (Nikon).…”