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영어

Recent technological progress in producing germline chimeras has provided an important means of studying genetic resource conservation, and transgenics production. Primordial germ cells (PGCs) are called gonadal germ cells (GGCs), after migration to the gonadal ridge. GGCs ultimately differentiate into spermatogonia in the testes or oogonia in the ovaries. Germline chimeras can be produced in domestic chicken by transferring GGCs collected from the embryonic gonad into the bloodstream of 2-dayold recipient embryos. However, it is diffcult to purify GGCs from the majority number of somatic cells after digesting embryonic gonads using proteinases such as trypsin and EDTA. The aim of the present study was to develop a simple method for isolating GGCs from the gonads of 7-day-old chick embryos. A simple method for isolating viable GGCs was developed in the domestic chicken. Developing gonads were isolated from 7-day-old chick embryos and cultured for up to 24 hours at 38.5℃ in phosphate buffered saline without Ca+ and Mg+ (PBS [-]). A discharge of GGCs from the gonad was observed within 40 minutes after introducing the embryonic gonad into PBS [-], and the number of discharged GGCs increased until 12 hours of incubation. The purity of the GGCs (number of discharged GGCs/total number of discharged cells) was approximately 48% for the initial 1 hours of incubation and decreased thereafter. These results indicate that high-purity, viable GGCs can be collected by simply introducing isolated developing gonads into PBS [-]. The results provide an alternative means for producing germ line chimeras to conserve genetic resources and to produce transgenic avian species.