DEVELOPMENT OF COST EFFECTIVE MICROTOOLS FOR OOCYTE ENUCLEATION DURING SCNT EMBRYO PRODUCTION

摘要

The present investigation was undertaken for developing a cost effective simplified tools and process of enucleation and electrofusion. For enucleation of matured goat oocytes, a bisection blade was developed by glueing a triangular sharp metallic blade with glass capillary (Fig. 1A), for cutting the zona digested ooplasm, having polar body and Mil chromosomes. The nucleated oocyte halve was instantly discarded by aspiration pipette (Fig. IB). The whole process was conducted using micromanipulator (Fig. 1C). Further, the process of electrofusion of ooplasm with cell was simplified for individual fusion of oocyte and somatic cell. For achieving this step, the L shaped Gentrode (BTX- Harvard, USA) was modified by developing innovative electrofusion system and process (Fig. 2A, 2B & 2C). which gave more than 80% electrofusion. The innovative enucleation and electrofusion tools and process adopted in our laboratory proved to be very efficient and cost effective technique.