Recombinant human activin A promotes development of bovine somatic cell nuclear transfer embryos matured in vitro
Hua, S.,Northwest A and F Univ., Yangling (China). Coll. of Veterinary Medicine | Lan, J.,Northwest A and F Univ., Yangling (China). Coll. of Veterinary Medicine | Liu, Y.G.,Northwest A and F Univ., Yangling (China). Coll. of Veterinary Medicine | Song, Y.L.,Northwest A and F Univ., Yangling (China). Coll. of Veterinary Medicine | Liu, J.,Northwest A and F Univ., Yangling (China). Coll. of Veterinary Medicine | Wang, Y.S.,Northwest A and F Univ., Yangling (China). Coll. of Veterinary Medicine | Zhang, T.,Northwest A and F Univ., Yangling (China). Coll. of Veterinary Medicine | Zhang, Y.,Northwest A and F Univ., Yangling (China). Coll. of Veterinary Medicine
To improve the culture system of bovine somatic cell nuclear transfer (SCNT) embryos, we studied the effects of activin A on developmental competence of bovine SCNT embryos during the early development stage based on the traditional culture method, and analyzed the expression level of the genes related to blastocyst hatching (Na/K-ATPase, Glut-1) and related to activin A signalling pathway (ActRII and Smad2). We generated the bovine SCNT embryo using a Holstein cow oocyte as recipient cytoplasm and a foetal ear fibroblast (Holstein cow, 120 days) as donor cell. The embryos were cultured as follows: experiment 1, the addition of activin A at the concentrations of 0 (control), 20 (M1-20), 40 (M1-40) or 80 ng/mL (M1-80) to the media during the first three days and no addition during the subsequent 5 days; experiment 2, no addition of activin A to the media during the first 3 days and the addition of activin A at the concentrations of 0 (control), 20 (M2-20), 40 (M2-40) or 80 ng/mL (M2-80) during the subsequent 5 days. The results indicated that the blastocyst formation rate and hatching rate, and total blastomere numbers as well as ICM/TE obtained in experiment 1 were not significantly different from the control group. In contrast, these values obtained in experiment 2 were significantly higher than in the control group. In addition, the relative abundance (ratio to GAPDH mRNA) of each gene (Glut-1, ActR II and Smad2) was not significantly different among the treatments in the experiment. The expression levels of 4 genes (Na/K-ATPase, Glut-1, ActR II and Smad2) in blastocysts obtained in experiment 2 were higher than those obtained in experiment 1. In conclusion, the present study suggests that the addition of activin A to the culture media from day 4 to day 8 can enhance the developmental competence of bovine SCNT embryos.
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