WCRB2014 POSTER PRESENTATIONS (1) (335 abstracts)
Steps forward canine primordial germ cell isolation and characterization
Aline Fernanda de Souza 1 , Naira Caroline Godoy Pieri 1 , Juliana Barbosa Cassals 1 , Ana Carolina Furlanetto Mançanares 1 , Kelly Cristine Santos Roballo 1 , Carlos Eduardo Ambrosio 2 , Felipe Perecin 1 & Daniele Santos Martins 1
1University of São Paulo, Pirassununga, Brazil; 2School of Veterinary Medicine and Animal Science, University of Sao Paulo, Sao Paulo, Brazil.
Introduction: Primordial germ cells (PGCs) are specified during early mammalian post implantation development. They are precursors of adult animals gametes and are considered stem cells of germline. Most of PGCs studies were performed in rodents or human with limited studies in other mammals such as canine. We have recently demonstrated a model to derive embryonic germ cells (EGC) in canine species.
Materials and methods: We isolated canine embryos between 25 and 30 days old and cultured on a continuous cell line of canine embryonal fibroblasts as feeder-cells in the presence of leukemia inhibitory factor (LIF) and basic fibroblast growth factor (bFGF). The culture characterization was detect using alkaline phosphatase activity. Immunophenotype of PGCs was evaluated by immunohistochemical with specific markers (Oct3/4, c-kit and Vasa).
Results and discussion: PGC in vitro culture from both pooled and individual embryos resulted in successful derivation of putative EGC and showed colony formation in early passages. Canine PGCs in culture showed similar morphology pluripotent stem cells. The EGCs expressed alkaline phosphatase activity, this result suggests a characteristic of pluripotency and germ cells in canine EGCs. Immunohistochemical analysis revealed Oct4/3, c-kit and Vasa expressions PGCs in the hind gut epithelium, in the mesenchyme of dorsal mesentery and in gonadal ridge development PGCs are cells expressing Oct3/4 after posgastrulation, which has a role in the totipotent phenotype while c-kit and Vasa controls proliferation, survival and migration.
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ISSN 2052-1472 (online)
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