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22 May 2014 An Efficient System to Establish Biopsy-Derived Trophoblastic Cell Lines from Bovine Embryos
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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.

Priscila Ramos-Ibeas, Alexandra Calle, Eva Pericuesta, Ricardo Laguna-Barraza, Rommel Moros-Mora, Ricaurte Lopera-Vásquez, Verónica Maillo, María Yáñez-Mó, Alfonso Gutiérrez-Adán, Dimitrios Rizos, and Miguel Ángel Ramírez "An Efficient System to Establish Biopsy-Derived Trophoblastic Cell Lines from Bovine Embryos," Biology of Reproduction 91(1), (22 May 2014).
Received: 8 February 2014; Accepted: 1 April 2014; Published: 22 May 2014

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