HTR8/SVneo Cells Display Trophoblast Progenitor Cell-Like Characteristics Indicative of Self-Renewal, Repopulation Activity, and Expression of “Stemness-” Associated Transcription Factors

Abstract

Introduction. JEG3 is a choriocarcinoma—and HTR8/SVneo a transformed extravillous trophoblast—cell line often used to model the physiologically invasive extravillous trophoblast. Past studies suggest that these cell lines possess some stem or progenitor cell characteristics. Aim was to study whether these cells fulfill minimum criteria used to identify stem-like (progenitor) cells. In summary, we found that the expression profile of HTR8/SVneo (CDX2+, NOTCH1+, SOX2+, NANOG+, and OCT-) is distinct from JEG3 (CDX2+ and NOTCH1+) as seen only in human-serum blocked immunocytochemistry. This correlates with HTR8/SVneo's self-renewal capacities, as made visible via spheroid formation and multi-passagability in hanging drops protocols paralleling those used to maintain embryoid bodies. JEG3 displayed only low propensity to form and reform spheroids. HTR8/SVneo spheroids migrated to cover and seemingly repopulate human chorionic villi during confrontation cultures with placental explants in hanging drops. We conclude that HTR8/SVneo spheroid cells possess progenitor cell traits that are probably attained through corruption of “stemness-” associated transcription factor networks. Furthermore, trophoblastic cells are highly prone to unspecific binding, which is resistant to conventional blocking methods, but which can be alleviated through blockage with human serum.

Protective effect of (±)α-tocopherol on brominated diphenyl ether-47-stimulated prostaglandin pathways in human extravillous trophoblasts in vitro

“…This work was performed with the HTR-8/SVneo cell line (Graham et al, 1993). The HTR-8/SVneo cell line was derived from first trimester placentae and has provided a useful cell culture model for studies of EVT cellular responses (Liu et al , 2012; Wang et al , 2012; Weber et al , 2013) and initial investigations of toxicant actions on EVTs (Park et al , 2014a; Tetz et al , 2013a). …”

Chemotherapy curable malignancies and cancer stem cells: a biological review and hypothesis

“…The model of cancer stem cells is perhaps less relevant to these cells and their malignancies as they arise from the initial cells that are formed at fertilisation and nuclear fusion. Whilst in this situation there can be no hierarchical cancer stem cells, as these cells arise from the first point in embryological development, recent work has indicated the presence of cancer cells with stem like properties developing within trophoblast cell lines [64]. We would suggest that the cells with stem like properties in this malignancy will share the innate sensitivity to chemotherapy that the trophoblast cells have naturally due to their close temporal relationship with the unique genetic event of nuclear fusion and the associated apoptotic sensitivity [4].…”

An international network (PlaNet) to evaluate a human placental testing platform for chemicals safety testing in pregnancy

“…hCG) is maintained in spheroid cultures, which can be dysregulated by transcription and mitosis inhibitors. Spheroids stained with tracer dye can also be assessed for their capacity to invade fresh placental explants within the hanging drop, providing functional data related to their "stemness" [86].…”

“…Preliminary attempts to micro-fabricate the cellular environment of the placenta, either in a multi-layered system to replicate placental barrier function, or as an organoid culture have shown promising potential, but these models require further characterisation with respect to toxicology testing and need to be better coupled to real-time analysis [84][85][86]. Advances in this field would facilitate xenobiotic testing, inclusive of more simplified placental transfer experiments and toxicology studies looking at effects on cell survival, homeostatic regulation and endocrinology.…”