The production of transgenic pigs for xenotransplantation

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1) Basic technoloeies for the production of transgenic pigs, such as in vitro maturation and fertilization and culture of oocytes, nuclear transfer, freezing of embryos, embryo transfer, have been established.2) mitosen-activated protein kinase (MAPK) mediates the maturation-inducing signal from the cytoplasm into nucleus and induces meiosis reinitiation in porcine oocytes.3) The effect of hyaluronic acid on the development of in vitro-produced porcine embryos varies with the condition of oocyte maturation and fertilization.4) A porcine cell line can be established from the hatched blastocyst and maintained in vitro for a long period, and that reconstructed embryos obtained by transferring the blastocyst-derived cells into enucleated oocytes have the ability to develop to the blastocyst stage in vitro.5) Vector including a gene encoding green fluorescent protein (GFP gene) was developed, and techniques for selection of the cells with GFP gene was established .6) Techniques of nuclear transfer of the cells with GFP gene into enucleated oocytes was established.