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Separation and in vitro culture of goat fetal testicular cells and behaviour of male germ-line stem cells (mGSCs)
Published online by Cambridge University Press: 13 June 2008
Abstract
Seminiferous tubule tissues from goat fetal testis were treated using four methods. One of the treatments, with 0.1% collagenase for 15 min and then with an enzyme mixture (containing 0.25% trypsin, 10 μg/ml DNase I, 0.04% EDTA) for 5 min showed ideal cell dispersal after centrifugation and washing. The in vitro culture of the cells separated with a different medium system showed that mulberry-shaped male germ-line stem cell (mGSC) clusters and a single layer of Sertoli cells appeared as the original generation in 120 h. The mGSC clusters developed in suspension and were distributed in different locations from the Sertoli cells in the plate. The cells in the marginal area of incompletely digested mGSCs clusters, and the single cells from them, were obviously homogenized after culture for 5 days in a fibre cell feeding layer from fetal mice (MEF feeding layer), whereas identically treated mGSCs clusters co-cultured with Sertoli cells in the same culture system did not displayed clear homogenization in marginal cells of the clusters. Also, the first generation of mGSC clusters co-cultured with Sertoli cells could form the same morphological properties with the original generation; cells in clusters were tight and mGSCs divided more slowly than those in the MEF feeding layer.
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