Purified gonocytes from the neonatal rat form foci of proliferating germ cells in vitro.
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-HSP90alpha (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.
Moore
T J
TJ
Department of Biochemistry, Cell Biology and Histology, Faculty of Veterinary Medicine, Utrecht University, 3508 TD Utrecht, The Netherlands.
de Boer-Brouwer
M
M
van Dissel-Emiliani
F M F
FM
D000818:Animals; D000831:Animals, Newborn; D000951:Antigens, Neoplasm / Q000032:analysis*; D015815:Cell Adhesion Molecules / Q000032:analysis*; D002455:Cell Division; D002469:Cell Separation / Q000379:methods*; D002478:Cells, Cultured; D000071858:Epithelial Cell Adhesion Molecule; D005434:Flow Cytometry; D008297:Male; D051381:Rats; D017208:Rats, Wistar; D013094:Spermatozoa / Q000502:physiology*
D000951:Antigens, Neoplasm; D015815:Cell Adhesion Molecules; D000071858:Epithelial Cell Adhesion Molecule