• Proceedings of the Korea Society of Poultry Science Conference
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• 2000.11a
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• pp.88-90
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• 2000

Although primordial germ cells(PGCs) have been used in the production of germline chimera, efficiency has not been satisfactory. The Present study was conducted to improve efficiency of germline chimera production using the cultured gonadal PGCs(gPGCs). Germline chimeric chickens were produced by transfer of cultured gonadal primordial germ cells from Korean Ogol Chicken (KOC) to White Leghorn (5.5-day-old) and cultured in vitro for 10 days. Approximately 200 gPGCs (2-day-old) recipient embryos from which blood had been withdrawn via the dorsal aorta prior to the injection. Recipient embryos were incubated until hatching. Germline chimerism of the chickens reaching maturity was examined by mating them with Korean Ogol Chicken. Donor-derived offspring were identified as germline chimeric chickens based on their feather color. The frequency of germline transmission of donor PGCs ranged 1.9∼60.7%. There was no difference between both sexes. Therefore, it can be concluded that efficiency of germline chimerism can be improved via using cultured gPGCs.

• Proceedings of the Korea Society of Poultry Science Conference
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• 2005.11a
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• pp.68-69
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• 2005

To identify germline chimeric chicken using germ cell transplantation method, the testcross, spends much time, labor and cost to perform, is the only way for distinguishing germline chimeric chicken from normal one And to enhance the method, development of breed-specific molecular markers have been needed. We have just identified breed-specific sequence polymorphisms among Barred Plymouth rock, Korean Ogol Chicken and White Leghorn in PMEL17 and MC1R gene the loci of which are identical to dominant white and extended black loci. These sequence polymorphism will be very useful for screening germline chimera.

• Development and Reproduction
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• v.11 no.3
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• pp.219-226
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• 2007

This study was conducted to identify optimistic primordial germ cells'(PGCs) migration activity using heat activated busulfan treatment for the increasing germline chimerism. Donar PGCs viability tests of important conditions for useful germ line chimerism indicated approximately $70{\sim}80%$ viability was time dependent. Transplantation experiments of PGCs into recipient embryos after busulfun treatment, showed the treatment group having 23.5% viability. By comparison, the control group showed 4.8% viability. The 96 hour treatment group and the 118 hour treatment group of the cultured PGCs showed high migration activity. Generally, the transplantation method would consider morphological and physiological characteristics before transplantation. In the present study, the effect of busulfan on migration activity showed viability highest at 53.4% after 48-hour incubation time. However, a previous study showed the best condition for transplantation time to be prior to the 48-hour incubation period, when the chicken embryo does not yet have a developed blood vessel system. In conclusion, an important condition for the production of a transgenic chicken is that most donor PGCs migrate into the recipient embryo without any inhibitory factors. The present results suggest, perhaps by using this modified method of transplantation, it can produce a more efficient chimeric germ line, transgenic chicken.

• Proceedings of the Korea Society of Poultry Science Conference
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• 2001.11a
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• pp.40-46
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• 2001

The use of pluripotent stem cells has tremendous advantages for various purposes but these cell lines with proven germ-line transmission have been completely established only in the mouse. Embryonic germ (EG) cell lines are also pluripotent and undifferentiated stem cells established from primordial germ cells (PGCs). This study was conducted to establish and characterize the chicken EG cells derived from gonadal primordial germ cells. We isolated gonadal PGCs from 5.5-day-old (stage 28) White leghorn (WL) embryos and established chicken EG cells lines with EG culture medium supplemented with human stem cell factor (hSCF), murine leukemia inhibitory factor (mLIF), bovine basic fibroblast growth factor (bFGF), human interleukin-11 (hIL-11), and human insulin-like growth factor-I (hIGF-I). These cells grew continuously for 4 months (10 passages) on a feeder layer of mitotically active chicken embryonic fibroblasts. These cells were characterized by screening with the Periodic acid-Shiff＇s reaction, anti-SSEA-1 antibody, and a proliferation assay after several passages. As the results, the chicken EG cells maintained characteristics of undifferentiated stem cells as well as that of gonadal PGCs. When cultured in suspension, the chicken EG cells successfully formed an embryoid body and differentiated into a variety of cell types when re-seeded onto culture dish. The chicken EG cells were injected into blastodermal layer at stage X and dorsal aorta of recipient embryo at stage 14 (incubation of 53hrs) and produced chimeric chickens with various differentiated tissues derived from the EG cells. The germline chimeras were also successfully induced by using EG cells. Thus, Chicken EG cells will be useful for the production of transgenic chickena and for studies of germ cell differentiation and genomic imprinting.