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2003•Chinese Journal of Coll BiologyRequires access

ISOLATION AND CULTURE OF PRIMORDIAL GERM CELLS FROM CHICKEN EMBRYOS

Xiao Xiao

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Abstract

Along with two methods of Ficoll density-gradient centrifugation and EDTA-Trypsin collection of isolating primordial germ cells (PGCs) from chicken embryos at three different stages: from blood at stage 14, from genital ridge at stage 19 and gonadal at stage 28. PGCs were then cultured in the TCM-199 medium containing 10% FCS in vitro. The number of viable cells were determined by using the Trypan Blue test; PGCs were identified using PAS reaction. The result showed that, EDTA-Trypsin collection was more preferable to PGCs collection at stage 19 and stage 28, and these two stages were suitable for collection of PGCs from early chicken embryos.

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What this paper is about

Along with two methods of Ficoll density-gradient centrifugation and EDTA-Trypsin collection of isolating primordial germ cells (PGCs) from chicken embryos at three different stages: from blood at stage 14, from genital ridge at stage 19 and gonadal at stage 28. PGCs were then cultured in the TCM-199 medium containing 10% FCS in vitro. The number of viable cells were determined by using the Trypan Blue test; PGCs were identified using PAS reaction. The result showed that, EDTA-Trypsin collection was more preferable to PGCs collection at stage 19 and stage 28, and these two stages were suitable for collection of PGCs from early chicken embryos.

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Available abstract

Along with two methods of Ficoll density-gradient centrifugation and EDTA-Trypsin collection of isolating primordial germ cells (PGCs) from chicken embryos at three different stages: from blood at stage 14, from genital ridge at stage 19 and gonadal at stage 28. PGCs were then cultured in the TCM-199 medium containing 10% FCS in vitro. The number of viable cells were determined by using the Trypan Blue test; PGCs were identified using PAS reaction. The result showed that, EDTA-Trypsin collection was more preferable to PGCs collection at stage 19 and stage 28, and these two stages were suitable for collection of PGCs from early chicken embryos.

Key concepts: Biology, Gonadal ridge, Trypan blue, Ficoll, Andrology, Embryo, Trypsin, Differential centrifugation

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