MicroRNA characterization in equine induced pluripotent stem cells

• DOI: 10.1371/journal.pone.0207074
• Publication Date: 2018-12-03T13:25:22Z
• Authors: Lucia Natalia Moro; Guadalupe Amin; Veronica Furmento; Ariel Waisman; Ximena Garate; Gabriel Neiman; Alejandro La Greca; Natalia Lucia San...; Carlos Luzzani; Gustavo E. Sevlever; Gabriel Vichera; Santiago Gabriel ...
• Keywords: Nuclear Transfer Techniques; Cancer Research; Cell biology; MicroRNA Regulation in Cancer and Development; Epigenetic Remodeling; Science; Induced Pluripotent Stem Cells; Gene; pluripotent; Kruppel-Like Factor 4; 03 medical and health sciences; stem cells; Biochemistry, Genetics and Molecular Biology; https://purl.org/becyt/ford/3.1; Health Sciences; Genetics; Animals; https://purl.org/becyt/ford/3; Horses; Molecular Biology; Biology; 0303 health sciences; Stem cell; microRNA; Gene Expression Profiling; Induced pluripotent stem cell; Q; R; Life Sciences; Induction and Differentiation of Pluripotent Stem Cells; MicroRNA; Cell Differentiation; Fibroblasts; MicroRNAs; Embryonic stem cell; Multipotent Mesenchymal Stem Cells; FOS: Biological sciences; Medicine; Research Article

Cell reprogramming has been well described in mouse and human cells. The expression of specific microRNAs demonstrated to be essential for pluripotent maintenance cell differentiation, but not much information is available domestic species. We aim generate horse iPSCs, characterize them evaluate the different (miR-302a,b,c,d, miR-205, miR-145, miR-9, miR-96, miR-125b miR-296). Two equine iPSC lines (L2 L3) were characterized after fibroblasts with four Yamanaka's factors (OCT-4/SOX-2/c-MYC/KLF4). pluripotency both was assessed by phosphatase alkaline activity, OCT-4, NANOG REX1 RT-PCR, immunofluorescence SOX-2 c-MYC. In vitro differentiation embryo bodies (EBs) showed capacity iPSCs differentiate into ectodermal, endodermal mesodermal phenotypes. MicroRNA analyses resulted higher miR-302 family, miR-9 miR-96 L2 L3 vs. (p<0.05), as previously shown Moreover, downregulation miR-145 miR-205 observed. After EBs, observed EBs respect also induced only EB-L2. addition, silico alignments mRNA targets suggested ability family regulate cycle epithelial mesenchymal transition genes, neural determinant genes similarly humans. conclusion, we could obtain determine first time level