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Electronic Journal of Biotechnology
Universidad Católica de Valparaíso
ISSN: 0717-3458
Vol. 17, No. 5, 2014, pp. 211-216
Bioline Code: ej14035
Full paper language: English
Document type: Research Article
Document available free of charge

Electronic Journal of Biotechnology, Vol. 17, No. 5, 2014, pp. 211-216

 en Establishment of a pheasant ( Phasianus colchicus check for this species in other resources ) spermatogonial stem cell line for the production of interspecies germ line chimeras
Kim, Jeong Hyun; Sharma, Neelesh; Kim, Sung Woo; Sodhi, Simrinder Singh; Ghosh, Mrinmoy; Kim, Nam Eun; Mongre, Raj Kumar; Oh, Sung Jong & Jeong, Dong Kee

Abstract

Background: Spermatogonial stem cells (SSCs) are important for the production of interspecies germ line chimeras. The interspecies germ cell transfer technique has been suggested as a way to conserve endangered birds. Our objective was to develop a technique for restoring endangered birds by developing interspecies germ line chimeras between pheasant ( Phasianus colchicus check for this species in other resources ) and chicken ( Gallus gallus check for this species in other resources ) with SSCs.
Results: SSCs were isolated fromthe surgically removed testis of a pheasant. Growth conditions for pheasant SSCs were established by co-culturing STO (SIM mouse embryo-derived thioguanine and ouabain resistant) cells and pheasant SSCs. The colony-forming cells divided and proliferated stably to yield an established SSC line. Pheasant SSCs showed strong reactivity for GDNF family receptor alpha1 (GFRα1) marker. Finally, production of germline chimeras was attempted by transferring pheasant SSCs into recipient embryos. Although final embryo survival was 5.6% (20/354), the initial survival rate was 88% (312/354). To measure the percent transfer of donor SSC to gonads, the pheasant SSCs were labeled with PKH 26 fluorescent dye. We observed 30% donor cells and 9.48% c-kit/CD117-positive cells in the gonads of recipient chickens. Donor SSCs were thus stably engrafted in the recipient gonads.
Conclusions: This study showed that SSCs can be used as a tool for the conservation of endangered birds and the production of germline chimeras. Our findings yield insights into howwe may use the pheasant spermatogonial stem cell line for efficient production of interspecies germ line chimeras and ultimately, to the restoration of endangered birds.

Keywords
Endangered birds; In vitro culture; Recipient chicken embryos; SSC transplantation

 
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