DEVELOPMENT OF THE SYSTEM FOR CREATING A TRANSGENIC MINIPIG WITH THE THREE-DIMENSIONAL MONITORING SYSTEM

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Aim of the project is to establish an efficient system of creating transgenic minipigs for heterogenic organ transplantation. We invented three-dimensional (3D) rigid endoscopes for obtaining less-invasively minipig ova from ovarium without laparotomy and an ova-holding device having 5 x 5 micropocket matrix made of microporous glass plate. The device can suck ova into micropockets having diameter of 150 micrometer with solution circulating across the plate fixed in the holder housing. Ova can be cultured in the device under controlled environments of appriate temperature, pH, and gas contents. GFP marked-human gene can be introduced into the nucleus with the computer guided microinjector. All the procedures can be monitored with the 3 D microscope and recorded with a video-cassette recorder. The cultured GFP- marked embryo can be selectively picked up and introduced laparoscopically into female minipig uterus less-invasively with the 3 D rigid endoscope. This system can create several ten times more efficiently a trangenic minipig than the conventional method which needs much skill and more female minipig sacrification.