Introduction Of Genes Relating To Muscular Dystrophy Into Chimeric Chickens By Embryo Engineering

Introduction of Chicken Muscular Dystrophy Gene

Akira Fujiwara1)' 2), Makoto Mizutani1), Tamao Ono2), Hiroshi Kagami2)

11Nippon Institute for Biological Science, Laboratory Animal Research Station, Kobuchisawa, Yamanashi 408-0041, Japan; 2Shinshu University, 8304 Minamiminowa, Nagano 399-4598, Japan

Abstract: A novel system has been developed to introduce genes relating to muscular dystrophy to chimeric chickens. Fertilized eggs were obtained from New Hampshire chicken; NH-413 strain which have genes relating to Fukuyama type muscular dystrophy. Blastoderms were isolated from these embryos. Cells from the center of area pellucida were specifically isolated from the blastoderms. Excess yolk were removed by PBS and only pluripotent cells were obtained. These cells were used as donor. Recipient embryos were obtained from White Leghorn chicken; Line-M. Approximately 700 cells were removed from the center of the area pellucida of the recipient in stage X blastoderm. The donor cells were microinjected into the subgerminal cavity of the recipients' blastoderms. The manipulated embryos were cultured ex vivo until hatching. The generated chimeric chickens had the donor derived brown plumage in the down, suggesting that the cells containing muscular dystrophy were introduced into the chimeras. These chimeric chickens will be raised until sexual maturity. The chimeric chickens will be back-crossed to donor strain; the New Hampshire. In case, these individuals would be germline chimeras, the New Hampshire's offspring would be breed very rapidly. These should be one of the powerful strategies for breeding and regeneration of chickens for an experimental animal model.

Key words: avian; pluripotent cells; cell culture

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