Developmental Potency of Goat Embryos Produced by Intra Cytoplasmic Sperm Injection and In Vitro Fertilization

Abstract

Joygeswar Bori, Morang Munmi, Bori Bhumika

The objective of this study was to standardize novel intracytoplasmic sperm injection (ICSI) technique, production of goat embryos by ICSI, in vitro fertilization (IVF) and comparison of their developmental potency. Immobilized sperm from epididymal semen with breaking tail were injected to mature metaphase ii oocytes by sharp micropipette of 5 μm diameter, as well as in vitro fertilization was carried out. ICSI and IVF derived zygotes were cultured in RVCL-blast-BSA media after chemical activation with ionomycin, CHX and 6-dmap. Sham injection was used as a control group. Results suggest that immobilized sperm with breaking tail microinjected by sharp injection pipette after chemical activation is an efficient approach for goat embryo production by ICSI. Satisfactory survival rate (83.2%) was found by using sharp injection pipette of 5 μm diameter. Chemical activation with ionomycin, CHX and 6-dmap was found effective with activation rate of 63.33%. Developmental rate of 2 cell, 4-8 cell, 8-16 cell, morula, blastocyst in ICSI and IVF group was 74.44%, 63.33%, 44.44%, 23.33%, 14.44% and 65.16%, 57.42%, 49.03%, 38.71%, 23.23%, respectively. Upon comparison, significantly higher (p<0.05) percentage of cleavage was found in ICSI group than IVF (73.40±3.12% vs. 64.06±2.44%, respectively). However, morula and blastocyst percentage were significantly higher (p<0.05) in IVF group than ICSI (23.20±2.73% vs. 37.90±3.15% and 14.56±1.75% vs. 21.05±2.09%). It can be concluded that ICSI with sharp micropipette of 5 μm diameter can be applied to assisted reproduction for in vitro goat embryo production.

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