TY - JOUR
T1 - Derivation and characterization of sleeping beauty transposon-mediated porcine induced pluripotent stem cells
AU - Kues, Wilfried A.
AU - Herrmann, Doris
AU - Barg-Kues, Brigitte
AU - Haridoss, Srividiameena
AU - Nowak-Imialek, Monika
AU - Buchholz, Thomas
AU - Streeck, Miriam
AU - Grebe, Antonia
AU - Grabundzija, Ivana
AU - Merkert, Sylvia
AU - Martin, Ulrich
AU - Hall, Vanessa Jane
AU - Rasmussen, Mikkel Aabech
AU - Ivics, Zoltan
AU - Hyttel, Poul
AU - Niemann, Heiner
PY - 2013/1/1
Y1 - 2013/1/1
N2 - 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.
AB - 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.
KW - Animals
KW - Antigens, Differentiation
KW - Cell Transformation, Neoplastic
KW - Cells, Cultured
KW - Coculture Techniques
KW - DNA Transposable Elements
KW - Fibroblasts
KW - Green Fluorescent Proteins
KW - Induced Pluripotent Stem Cells
KW - Mice
KW - Mice, Nude
KW - Microscopy, Fluorescence
KW - Neurogenesis
KW - Octamer Transcription Factor-3
KW - Promoter Regions, Genetic
KW - SOXB1 Transcription Factors
KW - Sus scrofa
KW - Teratoma
KW - Transcriptome
KW - Transgenes
U2 - 10.1089/scd.2012.0382
DO - 10.1089/scd.2012.0382
M3 - Journal article
C2 - 22989381
SN - 1547-3287
VL - 22
SP - 124
EP - 135
JO - Stem Cells and Development
JF - Stem Cells and Development
IS - 1
ER -