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Stem Cells Vol. 24 No. 7 July 2006, pp. 1638 -1645
doi:10.1634/stemcells.2005-0491; www.StemCells.com
© 2006 AlphaMed Press

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EMBRYONIC STEM CELLS

Progress Toward the Culture and Transformation of Chicken Blastodermal Cells

Youliang Wang, Carrie F. Brooks, Steven A. Jones, Lyn K. Olliff, Michael Morgan, Gordon L. Speksnijder, Chuck Foley, Alex J. Harvey

AviGenics, Inc., Georgia BioBusiness Center, Athens, Georgia, USA

Key Words. Avian transgenesis • Blastodermal cell culture • Embryonic stem cells

Correspondence: Alex J. Harvey, Ph.D.,AviGenics, Inc., Georgia BioBusiness Center, 111 Riverbend Road, Athens, Georgia 30605, USA. Telephone: 706-227-1170; Fax: 706-227-2180; email: harvey{at}avigenics.com

Received October 4, 2005; accepted for publication March 20, 2006.

Chicken blastodermal cells can be cultured for short periods of time and retain the ability to contribute to somatic and germline tissues when injected into {gamma}-irradiated stage X embryos. Such a method has yet to yield a germline transgenic bird, in part due to the low rate of transgene integration into the avian genome. In addition, the short culture period precludes the identification and expansion of those cells that carry an integrated transgene. In this study, two methods were developed that produced blastodermal cells isolated from stage X Barred Plymouth Rock embryos bearing an integrated transgene. Addition of chick embryo extract to the culture medium enabled expansion of single colonies for multiple passages. Southern blot analysis indicated that the transgenes had integrated as a single copy in most of the clones. Cells from passaged, transgenic embryo cells were injected into irradiated stage X White Leghorn embryos, producing hatched chicks that bore the donor cells in their somatic tissues. Transgene sequences were detected in sperm DNA; however, breeding of chimeras did not result in germline transmission of the transgene, indicating that the contribution of the transgenic cells to the germline was either nonexistent or very low.






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