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GV Extract Highly Induce Pig Stem Like Cells from Differentiated Somatic Cells
- 언어ENG
- URLhttps://db.koreascholar.com/Article/Detail/213392
Genomic reprogramming factors in the GV cytoplasm improved cloning efficiency in mice through the pre‐exposure of somatic cell nuclei to a GV cytoplasmic extract prior to nuclear transfer. In this study, a pig GV oocyte extract (pGV extract) was developed. Treatment of pig fibroblasts with the pGV extract promoted colony formation after 2–3 weeks in culture, concomitant with the expression of stem cell markers (Oct‐4, Rex1, Nanog, Sox2) and repression of differentiated cell markers (CKAP2, NPR3 ). Using fibroblasts transfected with human Oct‐4 promoter‐driven enhanced green fluorescent protein (Oct4‐EGFP), pGV extract treatment induced the reactivation of the Oct‐4 promoter in Oct4 ‐ EGFP cells by 10 days post‐treatment. These transgenic donor cells were injected into 8‐cell embryos. Oct‐4 promoter activity was subsequently detected in most ICM cells of the host blastocyst. Interestingly, reconstructed embryos with pGV extract‐treated Oct4‐ EGFP fibroblast nuclei showed prolonged expression of Oct4 in the ICM of embryos. Additionally, the pGV extract promoted somatic cell reprogramming and cloned embryo development when assessed by measuring histone H3‐K9 hypomethylation, the expression of Oct4 and Nanog in blastocysts, and the production of increased numbers of high‐ quality blastocysts. Under specific culture conditions, pGV extract‐treated fibroblast cells differentiated into neuronal, pancreas, cardiac, and endothelial lineages that were confirmed by antibodies against specific marker proteins. These data provide evidence for the generation of stem‐like cells from differentiated somatic cells by treatment with GV oocyte extracts in pig.
