Improved preimplantation development of porcine somatic cell nuclear transfer embryos by caffine treatment

Abstract

This study examined the effcts of a caffine treatment to improve nuclear reprogramming in porcine cloned embryos. Embryonic development and the expression of genes related to pluripotency (POU5F1, SOX2, NANOG, and CDX2) were compared aftr caffine supplementation during manipulation at diffrent concentrations (0, 1.25, 2.5, and 5.0 mM) and aftr varying the delayed activation time (control, 1, 2, and 4 h) aftr fusion. Caffine added to media during manipulation produced a higher rate of development to blastocysts in the 1.25 mM group than in the other concentration groups (22.8% vs. 16.1%, 16.2%, and 19.2%; p < 0.05). When caffine was added during the 4 h delayed activation, the 1.25 mM caffine concentration produced a signifiantly higher rate of development than those in the other 4 h-activation-delayed caffine concentration groups (22.4% vs. 9.4%, 14.0%, and 11.1%; p < 0.05). On the other hand, no signifiant improvement over that in the control group was observed when caffine was supplemented during both the manipulation period and delayed activation period (16.0% vs. 15.2%), respectively. The levels ofPOU5F1, SOX2, and NANOG expression in blastocysts were signifiantly higher in the delayed activation caffine group (4 h, 1.25 mM) than in the control group (1 h, 0 mM; p < 0.05). In conclusion, a caffine treatment at 1.25 mM during delayed activation for 4 h can improve the preimplantation development of porcine somatic cell nuclear transfer embryos by activating nuclear reprogramming.