Handmade cloned bovine embryos, parthenogenesis and in vitro fertilization: a comparison

Abstract

<p>The handmade cloning and parthenogenesis as a control leads to the possibility of studying biological processes such as cell reprogramming, epigenetics, embryo genome activation and pre- and post-natal development. The objective of this study was to standardize the process the handmade cloning (HCM) and to compare the resultant embryo production with parthenogenesis and in vitro fertilization (IVF). The primary fibroblast culture was established from explants of ear, the manual cloning technique was standardized with this fibroblast and 10 processes to cloning, parthenogenesis and fertilization in vitro were performed, and the compared production and embryo quality was performed by an ANOVA. An in vitro culture of fibroblast cells was established with optimal characteristics, allowing the genetic material to be used in the process of cloning. The parthenote group without zona pellucida (ZP) exhibited a higher cleavage rate compared with the other groups of parthenotes with ZP and IVF embryo (p<0.05). The number of blastomeres was greater in the IVF (109.81±11.70) compared with the parthenote with ZP (73.73±7.09), without ZP (78.16±7.65) and clone (CL, 77.5±8.23) groups. Embryo production in the CL, ZP, without ZP and IVF groups was not significantly different (29.7, 37.6, 33.8 and 35.2% respectively). The HCM technique was successfully standardized in the laboratory. The resultant embryo production was similar between hand-made cloned embryos, parthenogenesis and in vitro fertilization. The findings of this work give rise to different routes for studying embryology and contribute to the optimization of this technique for commercial purposes.</p>