Establishment and comparison of human term placenta-derived trophoblast cells

We established an immortalized term placenta-derived trophoblast cell line and demonstrated functional and transcriptomic differences against chorion trophoblasts and BeWo cells. Cells were isolated from term placenta utilizing enzymatic digestion, modified Percoll gradient-density isolation, and immunomagnetic purification. The resulting HLA-ABC (-) placental trophoblast cells (PTC) were immortalized using SV40T transfection to generate immortalized human PTC cytotrophoblasts (hPTCCTB). Additionally, these cells were transformed into syncytiotrophoblasts (hPTCSTB) using forskolin. The hPTCCTB cells were compared with their pPTC origin, choriocarcinoma cells (BeWo), and a commercially available PTC cell line using total RNA sequencing followed by principal component analysis (PCA) to determine differences in transcriptome profiles. The hPTCCTB cells were further compared with their pPTC origin for cell-specific markers (immunocytochemistry) and function (NF-kB and p38MAPK activations) after treatment with LPS and cigarette smoke extract (oxidative stress inducer).