Derivation and Characterization of Sleeping Beauty Transposon-Mediated Porcine Induced Pluripotent Stem Cells

作者：Kues Wilfried A^*; Herrmann Doris; Barg Kues Brigitte; Haridoss Srividiameena; Nowak Imialek Monika; Buchholz Thomas; Streeck Miriam; Grebe Antonia; Grabundzija Ivana; Merkert Sylvia; Martin Ulrich; Hall Vanessa J; Rasmussen Mikkel A; Ivics Zoltan; Hyttel Poul; Niemann Heiner

来源：Stem Cells and Development, 2013, 22(1): 124-135.

DOI：10.1089/scd.2012.0382

The domestic pig is an important large animal model for preclinical testing of novel cell therapies. Recently, we produced pluripotency reporter pigs in which the Oct4 promoter drives expression of the enhanced green fluorescent protein (EGFP). Here, we reprogrammed Oct4-EGFP fibroblasts employing the nonviral Sleeping Beauty transposon system to deliver the reprogramming factors Oct4, Sox2, Klf4, and cMyc. Successful reprogramming to a pluripotent state was indicated by changes in cell morphology and reactivation of the Oct4-EGFP reporter. The transposon-reprogrammed induced pluripotent stem (iPS) cells showed long-term proliferation in vitro over >40 passages, expressed transcription factors typical of embryonic stem cells, including OCT4, NANOG, SOX2, REX1, ESRRB, DPPA5, and UTF1 and surface markers of pluripotency, including SSEA-1 and TRA-1-60. In vitro differentiation resulted in derivatives of the 3 germ layers. Upon injection of putative iPS cells under the skin of immunodeficient mice, we observed teratomas in 3 of 6 cases. These results form the basis for in-depth studies toward the derivation of porcine iPS cells, which hold great promise for preclinical testing of novel cell therapies in the pig model.

• 出版日期2013-1