Isolation, Characterization, and Adoptive Transfer of Cultured Chicken Primordial Germ Cells

We have established feeder-less lines of Primordial Germ Cells (PGCs) from two chicken lines divergently selected for sperm mobility phenotype.  PGCs are extra-embryonic stem cells that populate the embryonic gonad and differentiate to give rise to sperm or ova depending upon the gender. Various studies have demonstrated the use of modified PGCs for generation of transgenic organisms. In this study we culturedPGCs from 4 day old chicken embryo. PGCs were cultured in modified DMEM media (without feeder layers). DNA and RNA were isolated and analyzed for gender and PGC specific gene expression. We are now introducing male specific PGCs into busulfan treated recipient embryos to determine whether PGC genotype determines sperm mobility phenotype. We found 68% embryo survivability after busulfan treatment. But the hatchability was reduced to 12% after PGCs injection. Surviving chicks are being reared to adulthood for analysis of sperm mobility phenotype of the donor PGCs.