Isolation and characterization of a bovine blastocyst-derived trophoblastic cell line, BT-1: Development of a culture system in the absence of feedercell

Citation
A. Shimada et al., Isolation and characterization of a bovine blastocyst-derived trophoblastic cell line, BT-1: Development of a culture system in the absence of feedercell, PLACENTA, 22(7), 2001, pp. 652-662
Citations number
34
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Reproductive Medicine","da verificare
Journal title
PLACENTA
ISSN journal
0143-4004 → ACNP
Volume
22
Issue
7
Year of publication
2001
Pages
652 - 662
Database
ISI
SICI code
0143-4004(200108)22:7<652:IACOAB>2.0.ZU;2-D
Abstract
We established a trophoblastic cell line, bovine trophoblast-1 (BT-1), deri ved from in vitro matured and fertilized blastocyst. While several trophobl astic cell lines have been previously reported using feeder cell, BT-1 coul d be cultured in the absence of feeder cell. BT-1 was Cultured for more tha n 18 months (over 75 passage) in the absence of feeder cells, using bovine endometrial fibroblast-conditioned medium (fibroblast-conditioned medium). We found that the cell growth was accelerated in fibroblast-conditioned med ium, In bromodeoxyuridine incorporation analysis, BT-1 cells growth rate in fibroblast-conditioned medium was about two-fold higher than that in conve ntional medium. Furthermore, fibroblast-conditioned medium accelerated atta chment of BT-1 cells to culture dishes following plating. BT-1 showed epith elial morphology and expressed cytokeratin. During continuous culture, cell s accumulated fluid under the cell sheet and form dome-like structure that eventually transformed into free floating vesicles, Reverse transcription p olymerase chain reaction analysis and immunoblot analysis demonstrated that BT-1 cells expressed interferon-tau as well as placental lactogcn (PL). Im munofluorescence analysis demonstrated that a small number of cells were PL -positive, and these cells were binucleate. The BT-1 trophoblastic cell lin e could serve as a powerful model system for the study of trophoblast cell lineage and proliferation. (C) 2001 Harcourt Publishers Ltd.