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Derivation and characterization of rat iPS cells using a mouse STEMCCA reprogramming vector
Author(s) -
Rajanahalli Pavan,
Weiss Mark L
Publication year - 2013
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.27.1_supplement.lb53
Subject(s) - induced pluripotent stem cell , reprogramming , embryonic stem cell , homeobox protein nanog , klf4 , biology , microbiology and biotechnology , sox2 , regenerative medicine , transgene , stem cell , genetics , cell , gene
Rats are considered an important animal model system by having many similarities to humans in terms of physiological processes of the cardiovascular, metabolic and neurological systems. Induced pluripotent stem cell (iPSC) technology has great potential to advance in the field of regenerative medicine by generating patient‐specific stem cell for treatment or in vitro disease modeling. Here, we derived rat iPSC's from Fischer 344 embryonic fibroblasts using a mouse STEMCCA cre‐excisable reprogramming vector expressing Oct‐4, Klf4, Sox‐2 and c‐Myc. Once rat ESC‐like colonies were derived, we removed the transgene vector and demonstrated that one of the clones tested showed a normal karyotype by G‐banding. The iPSC colonies demonstrate a morphology that is similar to rat embryonic stem cells (ESCs), are positively stained for pluripotency markers such as alkaline phosphatase, Oct‐4, Nanog, Sox‐2 and SSEA‐1. The rat iPSCs have been maintained more than 27 passages. Using a custom‐designed microarray, we compared the gene expression profile of rat iPSCs to rat ESCs and there was good similarity between the two with the exception of Cdx2 expression which is expressed in rat ESCs. We plan to further characterize the rat iPSCs to determine whether they can undergo neural differentiation in vitro and whether the micro RNA expression of rat iPSCs is similar to rat ESCs.