Effect of sperm and oocyte factors on Intracytoplasmic Sperm injection (ICSI) performance in mice and goats / Nor Farizah Abdul Hamid

Selected sperm and oocyte factors related were evaluated in ICSI experiments involving mouse and goat species in the present study. The mouse oocyte collection was conducted through oviduct retrieval, while the goat oocytes were retrieved via LOPU technique. The effectiveness of sperm capacitation chemical treatment (Heparin and Theophylline) for 1 hour and sperm movement (Rapid and Slow) were identified in both species. For oocyte factors, the effects of post-hCG injection durations (13-15 hours and 16-18 hours) in mice were studied. LOPU cycles (OR1, OR2 and OR3) and IVM durations (18-21 hours and 22-25 hours) were investigated in goats. In Experiments 1, the effects of different sperm capacitation chemicals and sperm movement of mouse sperm were studied. Mouse ICSI-embryonic development at early stages (2- and 4-cell stage) were improved by using Heparin compared to Theophylline (64.67±1.69% versus 60.50±1.21% and 50.55±2.01% versus 45.09±1.60%, respectively). Both sperm movement factors had same potential to produce mouse ICSI-derived embryos. In Experiments 2, the effect of post-hCG injection duration in mouse oocytes on embryonic development rates were studied. Two different durations were significantly different (P<0.05) on all cleavage rates (2-, 4-, 8-cell and morula stage) whereby at 13-15 hours were higher than at 16-18 hours duration (67.95±1.06% versus 57.02±1.33%, 53.51±1.13% versus 41.90±1.98%, 39.40±1.01% versus 22.21±1.62%, and 14.83±1.09% versus 9.77±1.02%, respectively). Experiment 3 evaluated the effect of two different sperm capacitation chemicals (Heparin and Theophylline) and sperm movement (Rapid and Slow) of goat sperm were studied. Both sperm capacitation chemicals gave same potential of goat ICSI-embryo iv development (P>0.05). Rapid sperm movement gave better goat ICSI-embryonic development (4-, 8- cell and morula stage), whereby the respective values were 56.62±4.69% versus 41.80±4.49%, 39.51±4.70% versus 24.24±4.05% and 9.21±2.85% versus 2.24±0.94%. Experiment 4 investigated the effect of LOPU cycles on the yield of different oocyte grades (Grades A, B, C, D and E) from 16 donors goat. Lower number of Grades D and E (31 and 19, respectively) oocytes were obtained compared to Grades A, B and C. Higher number of Grade C oocytes were obtained compared to Grades A and B from LOPU technique (153 versus 106 and 91, respectively). OR1 gave better quantity and quality oocytes, followed by OR2 and OR3 (number of oocytes per ovary were 5.47±0.67, 3.94±0.44 and 3.09±0.50, respectively). In another sub-experiment, maturation rate of goat oocytes at 18 to 21 hours was higher than 22 to 25 hours of IVM duration (97.34±7.86% versus 89.01±2.41%, respectively). However, both IVM durations had the same potential in ICSI-derived embryo development. In conclusion, for mouse study, embryonic development can be achieved from Heparin-sperm capacitation treatment, using any sperm movement for 13 to 15 hours of post-hCG suntikan oocytes. For goat study, LOPU is good procedure to provide consistent high quality oocytes (Grades A, B and C) and it usage can be repeated on the same donor up to 3 times. Rapid sperm movement is a better choice for ICSI-embryonic development. Both sperm capacitation chemical treatments have the same potential to produce ICSI-embryonic development.