Document Detail

Growth requirements and chromosomal instability of induced pluripotent stem cells generated from adult canine fibroblasts.
MedLine Citation:
PMID:  23016947     Owner:  NLM     Status:  MEDLINE    
In mice and humans, it has been shown that embryonic and adult fibroblasts can be reprogrammed into pluripotency by introducing 4 transcription factors, Oct3/4, Klf4, Sox2, and c-Myc (OKSM). Here, we report the derivation of induced pluripotent stem cells (iPSCs) from adult canine fibroblasts by retroviral OKSM transduction. The isolated canine iPSCs (ciPSCs) were expanded in 3 different culture media [fibroblast growth factor 2 (FGF2), leukemia inhibitory factor (LIF), or FGF2 plus LIF]. Cells cultured in both FGF2 and LIF expressed pluripotency markers [POU5F1 (OCT4), SOX2, NANOG, and LIN28] and embryonic stem cell (ESC)-specific genes (PODXL, DPPA5, FGF5, REX1, and LAMP1) and showed strong levels of alkaline phosphatase expression. In vitro differentiation by formation of embryoid bodies and by directed differentiation generated cell derivatives of all 3 germ layers as confirmed by mRNA and protein expression. In vivo, the ciPSCs created solid tumors, which failed to reach epithelial structure formation, but expressed markers for all 3 germ layers. Array comparative genomic hybridization and chromosomal fluorescence in situ hybridization analyses revealed that while retroviral transduction per se did not result in significant DNA copy number imbalance, there was evidence for the emergence of low-level aneuploidy during prolonged culture or tumor formation. In summary, we were able to derive ciPSCs from adult fibroblasts by using 4 transcription factors. The isolated iPSCs have similar characteristics to ESCs from other species, but the exact cellular mechanisms behind their unique co-dependency on both FGF2 and LIF are still unknown.
Sehwon Koh; Rachael Thomas; Shengdar Tsai; Steve Bischoff; Ji-Hey Lim; Matthew Breen; Natasha J Olby; Jorge A Piedrahita
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2012-11-28
Journal Detail:
Title:  Stem cells and development     Volume:  22     ISSN:  1557-8534     ISO Abbreviation:  Stem Cells Dev.     Publication Date:  2013 Mar 
Date Detail:
Created Date:  2013-03-01     Completed Date:  2013-08-12     Revised Date:  2014-03-27    
Medline Journal Info:
Nlm Unique ID:  101197107     Medline TA:  Stem Cells Dev     Country:  United States    
Other Details:
Languages:  eng     Pagination:  951-63     Citation Subset:  IM    
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MeSH Terms
Alkaline Phosphatase / metabolism
Antigens, Differentiation / genetics,  metabolism
Cell Differentiation
Cell Shape
Cells, Cultured
Chromosomal Instability
Coculture Techniques
Comparative Genomic Hybridization
Fibroblasts / physiology*
Gene Expression
In Situ Hybridization, Fluorescence
Induced Pluripotent Stem Cells / enzymology,  pathology*,  transplantation
Mice, SCID
Recombinant Proteins / biosynthesis,  genetics
Teratoma / genetics,  pathology
Transcription Factors / biosynthesis,  genetics
Reg. No./Substance:
0/Antigens, Differentiation; 0/Recombinant Proteins; 0/Transcription Factors; EC Phosphatase

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