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A Testis-Mediated Germline Chimera Production Based on Transfer of Chicken Testicular Cells Directly into Heterologous Testes¹

Department of Food and Animal Biotechnology,³ Seoul National University, Seoul 151-921, Korea Bioresources Institute,⁴ EasyBio System, Chonan, Choongcheongnam-Do 330-822, Korea Avicore Biotechnology Institute Inc.,⁵ Gyeonggi-Do 435-824, Korea College of Bioresources Science,⁶ Dankuk University, Chonan, Choongcheongnam-Do 330-714, Korea

ABSTRACT

In this study, we proposed a testis-mediated germline chimera production system based on the transplantation of testicular cells directly into heterologous testes. The testicular cells of juvenile (4-wk-old) or adult (24-wk-old) Korean Ogol chickens with a recessive pigmentation inhibitory gene, with or without prior culture, were injected (2 x 10⁷ cells/head) into the seminiferous tubules of juvenile or adult recipients with White Leghorn with a dominant pigmentation inhibitory gene in a 2 x 2 factorial arrangement. The localization of transplanted cells into the inner space of the seminiferous tubules was confirmed within 24 h after injection. Subsequent testcross analyses showed that 7.8% (5/64) of the recipients had chimeric status in their testes. The periods of time from transfer to hatching of the first progeny with black feathers were 38 and 45 days for adult cells transplanted into an adult recipient, 188 days for adult cells into a juvenile recipient, and 137 days for juvenile cells into a juvenile recipient. Culture of the testicular cells derived both colony-forming and monolayer-forming cells. The colony-forming cells were stained positively for periodic acid Schiff solution, and further reacted with anti-SSEA-1, anti-SSEA-3, and anti-SSEA-4 antibodies both before and after culture for 15 days. In conclusion, it may be possible to develop the testis-mediated germline chimera production technique, which extends the feasibility of genetic manipulations in avian species.

chicken, chimera, gamete biology, germline, male reproductive tract, spermatogenesis, testicular cells, testis

¹ Supported by a grant from the BioGreen 21 program, Rural Development Administration, Korea and a graduate fellowship provided by the Korean Ministry of Education through the Brain Korea 21 project.

² Correspondence: FAX: 822 874 4811; jaehan{at}snu.ac.kr

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