Valproic acid assisted reprogramming of fibroblasts for generation of pluripotent stem cells in buffalo (Bubalus bubalis).

摘要: Generation of pluripotent stem cells by reprogramming somatic quality animals has numerous potential applications in agricultural and biomedical sciences. Unfortunately, till now, buffalo fetal fibroblast (bFFs) been very ineffient despite intensive efforts. Here, we attempted to enhance efficiency using the HDAC inhibitor valproic acid (VPA) bFFs transfected with pLentG-KOSM pseudo virus carrying mouse specific genes. FACS analysis revealed that VPA treatment significantly increased (p < 0.05) GFP+ comparison untreated control. Further, among different concentrations, 1.5 mM was found be optimal, increasing about 5 fold 2.5-fold colonies (P larger size as compared These were further propagated characterised. The displayed embryonic cell (ESC)-like morphology, normal karyotype, positive for alkaline phosphatase staining well immune-positive ESC markers Oct4, Nanog, SSEA1, TRA-1-60 TRA-1-81. primary higher expression genes than control, which declined gradually on subsequent passages. reprogrammed readily formed embryoid bodies vitro all three germ layers. results indicated viral transducted can improve generation induced help their long term maintenance buffalo.