title: Establishment and Characterization of a Telomerase-Immortalized Sheep Trophoblast Cell Line.
abstract: The primary sheep trophoblast cells (STCs) have a finite lifespan in culture. This feature limits the scope for long-term in vitro studies with STCs. This study was an attempt to establish and characterize a telomerase-immortalized sheep trophoblast cell line. STCs were isolated and purified by using Percoll and specific immunoaffinity purification, respectively. The purified STCs were transfected with a plasmid carrying sequences of human telomerase reverse transcriptase (hTERT) to create immortalized sheep trophoblast cell line (hTERT-STCs). hTERT-STCs showed a stable expression of hTERT gene, serially passaged for a year, and showed active proliferation without signs of senescence. Cytokeratin 7 (CK-7), secreted human chorionic gonadotrophin subunit β (CG-β), placental lactogen (PL), and endogenous jaagsiekte sheep retrovirus (enJSRV) envelope genes were expressed in hTERT-STCs. Transwell cell invasion assay indicated that hTERT-STCs still possessed the same invasive characteristics as normal primary sheep trophoblast cells. hTERT-STCs could not grow in soft agar and did not develop into tumors in nude mice. In this study, we established a strain of immortalized sheep trophoblast cell line which could be gainfully employed in the future as an experimental model to study trophoblast cells with secretory function, invasive features, and probable biological function of enJSRV envelope genes.
authors.affiliation: College of Veterinary Medicine, Inner Mongolia Agricultural University, Hohhot 010018, China.
authors.forename: Yufei
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: College of Veterinary Medicine, Inner Mongolia Agricultural University, Hohhot 010018, China.
authors.forename: Jing
authors.identifier: 0000-0002-9257-6060
authors.initials: J
authors.lastname: Shi
authors.affiliation: College of Veterinary Medicine, Inner Mongolia Agricultural University, Hohhot 010018, China; Key Laboratory of Clinical Diagnosis and Treatment Technology in Animal Disease, Ministry of Agriculture, Hohhot 010018, China.
authors.forename: Shuying
authors.initials: S
authors.lastname: Liu
mesh_terms: D000818:Animals; D002461:Cell Line, Transformed; D015870:Gene Expression; D006801:Humans; D012756:Sheep; D019098:Telomerase / Q000096:biosynthesis* / Q000235:genetics; D014327:Trophoblasts / Q000166:cytology* / Q000201:enzymology*
chemical_list: C509186:TERT protein, human; D019098:Telomerase