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KMID : 0900920000240040385
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Korean journal of Animal Reproduction
2000 Volume.24 No. 4 p.385 ~ p.394
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Characterization of Apoptosis in Porcine Primordial Germ Cells In Vitro
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Lee C.-K.
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Abstract
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When porcine primordial germ cells (PGCs) isolated from the genital ridge and placed in culture to establish EG cells, a large proportion of PGCs are lost during the early period of culture. To characterize the in vitro death of porcine PGCs, PGCs were cultured in suspension, and apoptosis analyzed using a fluorescent activated cell sorter-based DNA fragmentation assay. The results from flow cytometric analysis showed an increase in apoptosis in cultured cells. However, the cells isolated from the genital ridges are a mixture of PGCs and somatic cells. To detect apoptotic signals specific from porcine PGCs, quantitative TUNEL assay was performed at different time of culture (0 ∼ 24 h). The proportion of apoptotic porcine PGCs determined by double staining with alkaline phosphatase activity and in situ TUNEL assay increased as the time of culture progressed and continued at least 24 h. These results demonstrate that one of the causes of loss of porcine PGCs in vitro is apoptosis.
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KEYWORD
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Porcine POCs, Apoptosis, TUNEL assay
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