Abstract
Induced pluripotent stem cells (iPSCs) hold a great potential for therapeutic regenerative medicine. The aim of this study was to generate induced pluripotent stem cells from goat embryonic cardiac tissue derived fibroblasts. The isolated cardiac fibroblasts from the cardiac tissue of goat embryos were positive for alfa smooth muscle actin, vimentin and discoidin domain receptor2. From these cells, we generated transgene free iPSCs using piggyBac transposons / transposase using five transcription factors (Oct4, Sox2, Klf, Myc and Lin 28). The generated iPSCs were SSEA1, SSEA4 and Oct4 positive. They were cultured on neofeeders using 20% Serum replacement - IMDM with bFGF. They could form cystic and compact embryoid bodies that showed differentiated ectodermal and mesodermal like cells when cultured using 20% FBS-IMDM without bFGF. The iPSCs, generated in the frame of this approach were produced without the use of integrating virus and the reprogramming transgenes were removed at the end of the process. Though there were limitations in the approach used, a substantial sign of reprogramming was obtained.
Highlights
Generation of induced pluripotent stem cells (iPSCs) is a highly meticulous procedure which has been introduced into the field of regenerative medicine using different methods
Goat cardiac fibroblasts were isolated according to the protocol discussed in the materials and methods and cultured on gelatincoated cell culture dishes with GCF culture medium
GCFs were probed with antibodies for cardiac fibroblast markers αSMA, vimentin and Discoidin doman receptor 2 (DDR2) showing positive results (Figure 1B-D)
Summary
Generation of iPSCs is a highly meticulous procedure which has been introduced into the field of regenerative medicine using different methods. Prior to the use of iPSC-derived cells or tissues in clinical application in human patients; their safety, integration and lack of tumorigenicity has to be investigated in proper animal models bearing significant similarities to humans[4]. Examples of such studies include those that were done on equine[5], pigs[6] and cattle[7], each with a variable degree of success
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