Unlike mouse results, cloning efficiency of nuclear transfer from porcine induced pluripotent stem cells (piPSCs) is very low. The present study was performed to investigate the effect of cell cycle inhibitors on the cell cycle synchronization of piPSCs. piPSCs were generated using combination of six human transcriptional factors under stem cell culture condition. To examine the efficiency of cell cycle synchronization, piPSCs were cultured on a matrigel coated plate with stem cell media and they were treated with staurosporine (STA, 20 nM), daidzein (DAI, 100 μM), roscovitine (ROSC, 10 μM), or olomoucine (OLO, 200 μM) for 12 h. Flow Cytometry (FACs) data showed that piPSCs in control were in G1 (37.5±0.2%), S (34.0±0.6%) and G2/M (28.5±0.4%). The proportion of cells at G1 in DAI group was significantly higher than that in control, while STA, ROSC and OLO treatments could not block the cell cycle of piPSCs. Both of viability and apoptosis were affected by STA and ROSC treatment, but there were no significantly differences between control and DAI groups. Real-Time qPCR and FACs results revealed that DAI treatment did not affect the expression of pluripotent gene, Oct4. In case of OLO, it did not affect both of viability and apoptosis, but Oct4 expression was significantly decreased. Our results suggest that DAI could be used for synchronizing piPSCs at G1 stage and has any deleterious effect on survival and pluripotency sustaining of piPSCs.

• Seongsoo Hwang(Animal Biotechnology Division, National Institute of Animal Science) Corresponding Author
• Gi-Sun Im(Animal Biotechnology Division, National Institute of Animal Science, RDA)
• Jae-Seok Woo(Animal Biotechnology Division, National Institute of Animal Science, RDA)
• Keon Bong Oh(Animal Biotechnology Division, National Institute of Animal Science, RDA)
• Sun-A Ock(Animal Biotechnology Division, National Institute of Animal Science, RDA)
• Mi-Ryung Park(Animal Biotechnology Division, National Institute of Animal Science, RDA)
• Hyeon Yang(Animal Biotechnology Division, National Institute of Animal Science, RDA)
• Tae-Uk Kwak(Animal Biotechnology Division, National Institute of Animal Science, RDA)
• In-Sul Hwang(Animal Biotechnology Division, National Institute of Animal Science, RDA) ae-Jin Kwon and In-Sul Hwang have contributed equally to this work.
• Dae-Jin Kwon(Animal Biotechnology Division, National Institute of Animal Science, RDA, 2International Agricultural Development and Cooperation Center, Chonbuk National University) Dae-Jin Kwon and In-Sul Hwang have contributed equally to this work.