In vitro Culture of Testicular and Ovarian Gonocytes Obtained from 19-day Incubated Chicken Embryos and Subsequent Colonization into Gonads of Recipient Embryos

Testicular gonocytes differentiate into spermatogonia and spermatogonial stem cells could be established by culturing spermatogonia in vitro. Spermatogonial stem cells differentiate exclusively into spermatozoa and thus they are very useful for genetic manipulation in chickens. The present study was carried out to examine the possibility of in vitro proliferation of testicular and ovarian gonocytes in chickens. Gonads were obtained from 19-day incubated chicken embryos and the dissociated gonadal cells were cultured in vitro. Then, non-adherent cells containing gonocytes were collected and cultured further. It was confirmed that a part of the testicular gonocytes proliferated in vitro, when analyzed by anti-CVH antibody. The proliferated testicular gonocytes occasionally formed cell colonies. The cultured testicular gonocytes were successfully implanted in the gonads by transferring them into the coelomic epithelium that corresponds to the future gonadal region of recipient embryos. However, no apparent proliferation was observed in the ovarian gonocytes in vitro. These results suggest that testicular goncytes can be cultured in vitro and are one of the candidate cells for genetic manipulation in chickens.