WCRB2014 POSTER PRESENTATIONS (1) (335 abstracts)
Derivation of bovine induced pluripotent stem cells by a transposon approach
Talluri Thirumala Rao 1 , Dharmendra Kumar 2 , Silke Glage 3 , Wiebke Garrels 3 , Katharina Debowski 4 , Rudiger Behr 4 , Heiner Niemann 1 & Wilfried Kues 1
1Institute of Farm Animal Genetics, Neustadt am rbge, Germany; 2Central Institute for Research on Buffaloes, Hisar, India; 3Medical School Hannover, Hannover, Germany; 4German Primate Center, Göttingen, Germany.
Introduction: Induced pluripotent stem (iPS) cells are considered as a seminal breakthrough in stem cell research and are promising for the development of advanced regenerative therapies. Considering the potential of this technology for both basic and clinical research, it is tempting to extend this research to important livestock species, such as cattle, in which ES cell lines are yet not available.
Materials and methods: Here, we describe a non-viral method for the derivation of bovine iPS cells, employing the piggyBac (PB) transposon system. The reprogramming PB transposon encode the reprogramming factors OCT4, SOX2, KLF4, MYC, LIN28 and NANOG, each separated by self-cleaving peptide sequences and driven by the chimeric CAGGS promoter. One PB transposon-reprogrammed bovine iPS (biPS) cell line was established.
Results and discussion: One PB transposon-reprogrammed bovine iPS (biPS) cell line was established. The derived bovine iPS line expressed typical markers of embryonic stem cells (OCT4, SOX2, c-MYC, KLF4 and NANOG), showed long term proliferation under feeder-free culture conditions, differentiated into derivatives of the three germ layers in vitro, and formed teratomas after SUBCUTANEOUS injection into immune deficient nude mice. These results are a major step towards the derivation of authentic bovine iPS cells, in which the transposon transgenes can be eliminated after reprogramming.
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ISSN 2052-1472 (online)
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