WCRB2014 POSTER PRESENTATIONS (1) (335 abstracts)
Derivation of porcine iPS-like cells from fibroblast of a translocated azoospermic boar
Annabelle Congras 1 , Harmonie Barasc 2 , Florence Vignoles 1 , Chantal Delcros 1 , Alain Pinton 2 , Kamila Canale-Tabet 1 , Olivier Feraud 3 , Ali Turhan 4 , Martine Yerle-Bouissou 1 & Herve Acloque 1
1INRA, Castanet-Tolosan, France; 2ENVT, Toulouse, France; 3INSERM, Villejuif, France; 4INSERM, Poitiers, France.
Chromosomal rearrangements have a crucial impact on the proper proceedings of meiosis and can lead by several mechanisms to the production of unbalanced gametes or to the complete arrest of gametes production. To assess the impact of these rearrangement in the early development of pig germ cells, we proposed to generate a library of stem cells from an azoospermic boar carrying a reciprocal translocation t(Y:14), as a new tool for the development of an in vitro differentiation system from pluripotent stem cells to germ cells.
We report the reprogramming of translocated fibroblast by integrative or non-integrative viral overexpression of Oct4, Sox2, Klf4 and c-Myc. iPS cell lines were characterized for pluripotency, cell cycle and differentiation potential by conventional methods. Genomic stability was analyzed by G-banding karyotype, comparative genomic hybridization and FISH.
The porcine iPS-like cell lines harbored characteristics of ground and naïve pluripotency when cultured in specific media. They expressed several pluripotency genes and harbored an ES-like cell cycle. Nevertheless, contrary to mouse and human iPS, they did not silence the integrated exogenes, leading to a poor differentiation potential. Moreover, cytogenetic analysis revealed a high genomic instability upon passaging which suggest the development of population with an increased selective advantage. We characterized the selected duplications and compared them to those previously described in other species. In contrast, non-integrative reprogrammation system gives us promising results regarding differentiation potential and genomic stability and will bring new insights into the molecular factors controlling and maintaining pluripotency in the pig species.
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ISSN 2052-1472 (online)
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