Development of a modified method of handmade cloning in dromedary camel.

摘要: In this study, a modified method of handmade cloning (m-HMC), which had been originally developed in sheep, was used for somatic cell nuclear transfer (SCNT) the dromedary camel. The unique feature m-HMC over current SCNT methods lies use simple device (a finely drawn micropipette made Pasteur pipette) chemically-assisted enucleation oocytes under stereomicroscope with improved efficiency and ease operation. Using system, throughput cloned embryo reconstitution increased 2-fold compared to control (c-NT). Stepwise measurement reactive oxygen species (ROS) revealed that method, steps, duration all influenced oxidative activity oocytes, but their impact were not similar. Specifically, UV-assisted oocyte identified as major source ROS production, explained significantly higher total reconstituted embryos c-NT m-HMC. Fusion (95.3±3.3 vs. 75.4±7.6%) blastocyst development (22.5±3.0 14.1±4.3%) c-NT, respectively, blastocysts transferable quality obtained similar rates (41.9±8.2 48.0±15.2%, respectively). Significance differences observed number (155.3±13.6 123.6±19.5) trophectoderm (145±9.5 114.3±15.2), inner mass (10.3±4.1 9.3±5.3) counts between m-HMC, respectively. However, expression key developmental genes (POU5F1, KLF4, SOX2, MYC, CDX2) comparable both groups. introduced might be viable approach efficient production camel clones research commercial utilization.