A study in development of transgenic cattles for the use of hybrid bioartifical liver.

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In research on extracorporeal bioartificial liver perfusion system using xenogeneic liver, the chief purpose of the transgenic technique is to knock out genes of xenogeneic proteins, and transduce genes of human proteins into the animal. As an initial approach, we planned to transduce human albumin cDNA into porcine germ cells to produce porcine liver which can secret human albumin for the purpose of utilizing it for bioartificial liver support, we produced a gene of human albumin cDNA (hAlb) and EGFP (enhanced green fluorescent protein) gene, pCX-hAlb-EGFP. We transfected it initially into NIH3T3 cells using lipofection method, secondly into mouse germ cells by classic microinjection method, and thirdly into porcine germ cells by intracytoplasmic sperm injection (ICSI) method in order to induce expression of human albumin in the cultured cells and animal liver. As the results, NIH3T3 cells showed ultraviolet lightening and expressed human albumin in Northern hybridization analysis, and a transgenic pig was produced of which the skin showed ultraviolet lightening and integration of human albumin gene in PCR analysis. It was a pity that the pig was born dead, but this was initial epoch-making report of expression of EGFP and human albumin in TG pig produced by ICSI method.