title: An efficient system to establish biopsy-derived trophoblastic cell lines from bovine embryos.
abstract: Trophoblastic cells play a crucial role in implantation and placentogenesis and can be used as a model to provide substantial information on the peri-implantation period. Unfortunately, there are few cell lines for this purpose in cattle because of the difficulty of raising successive cell stocks in the long-term. Our results show that the combination of a monolayer culture system in microdrops on a surface treated with gelatin and the employment of conditioned media from mouse embryonic fibroblasts support the growth of bovine trophoblastic cells lines from an embryo biopsy. Expression profiles of mononucleate- and binucleate-specific genes in established trophoblastic cells lines represented various stages of gestation. Moreover, the ability to expand trophoblastic cell lines for more than 2 yr together with pluripotency-related gene expression patterns revealed certain self-renewal capacity. In summary, we have developed a system to expand in vitro trophoblastic cells from an embryo biopsy that solves the limitations of using amplified DNA from a small number of cells for bovine embryo genotyping and epigenotyping and, on the other hand, facilitates the establishment of trophoblastic cell lines that can be useful as peri-implantation in vitro models.
authors.affiliation: Departamento de Reproduccion Animal, INIA, Madrid, Spain.
authors.forename: Priscila
authors.initials: P
authors.lastname: Ramos-Ibeas
authors.affiliation: Departamento de Reproduccion Animal, INIA, Madrid, Spain.
authors.forename: Alexandra
authors.initials: A
authors.lastname: Calle
authors.affiliation: Departamento de Reproduccion Animal, INIA, Madrid, Spain.
authors.forename: Eva
authors.initials: E
authors.lastname: Pericuesta
authors.affiliation: Departamento de Reproduccion Animal, INIA, Madrid, Spain.
authors.forename: Ricardo
authors.initials: R
authors.lastname: Laguna-Barraza
authors.affiliation: Departamento de Reproduccion Animal, INIA, Madrid, Spain.
authors.forename: Rommel
authors.initials: R
authors.lastname: Moros-Mora
authors.affiliation: Departamento de Reproduccion Animal, INIA, Madrid, Spain.
authors.forename: Ricaurte
authors.initials: R
authors.lastname: Lopera-Vásquez
authors.affiliation: Departamento de Reproduccion Animal, INIA, Madrid, Spain.
authors.forename: Verónica
authors.initials: V
authors.lastname: Maillo
authors.affiliation: Hospital Universitario Santa Cristina, Instituto de Investigaciones Sanitarias Princesa, Madrid, Spain.
authors.forename: María
authors.initials: M
authors.lastname: Yáñez-Mó
authors.affiliation: Departamento de Reproduccion Animal, INIA, Madrid, Spain.
authors.forename: Alfonso
authors.initials: A
authors.lastname: Gutiérrez-Adán
authors.affiliation: Departamento de Reproduccion Animal, INIA, Madrid, Spain.
authors.forename: Dimitrios
authors.initials: D
authors.lastname: Rizos
authors.affiliation: Departamento de Reproduccion Animal, INIA, Madrid, Spain ramirez@gmail.com.
authors.forename: Miguel Ángel
authors.initials: MÁ
authors.lastname: Ramírez
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000166:cytology*; D002417:Cattle; D018929:Cell Culture Techniques; D002460:Cell Line / Q000166:cytology*; D010064:Embryo Implantation; D004622:Embryo, Mammalian / Q000166:cytology*; D015870:Gene Expression; D014327:Trophoblasts / Q000166:cytology*
keywords: bovine; camel; conditioned media; embryo biopsy; embryo culture; ruminants; trophoblast; trophoblastic cell lines