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Purified Gonocytes from the Neonatal Rat form Foci of Proliferating Germ Cells in Vitro

Department of Biochemistry, Cell Biology and Histology, Faculty of Veterinary Medicine, Utrecht University, 3508 TD, Utrecht, The Netherlands

Abstract

Due to the lack of a specific marker for gonocytes from newborn rats, isolation of these cells has proven difficult and laborious. We have found a specific cell membrane marker, the Epithelial Cellular Adhesion Molecular (Ep-CAM) that can be used to isolate these cells using antibody directed cell sorting. 4 days post partum (dpp) rat testes were enzyme treated to attain a cell suspension, which was labelled with an antibody (GZ1) against Ep-CAM and tagged with a fluorescent probe. The labelled cell suspension was run through a FACS cell sorter, from which a gonocyte suspension of >85% purity was attained. The cells remained viable in culture and proliferated actively as determined by double labelling the cells with anti-HSP90 (a specific germ cell marker) and anti-BrdU antibodies (after BrdU incorporation). During culture, these cells formed chains of 2 to 4 cells and aggregates of proliferating germ cells were found after 8 days of culture.

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