Purified gonocytes from the neonatal rat form foci of proliferating germ cells in vitro

T J Moore, M de Boer-Brouwer, F M F van Dissel-Emiliani

Research output: Contribution to journalArticlepeer-review

19 Citations (Scopus)

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-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.

Original languageEnglish
Pages (from-to)3171-4
Number of pages4
JournalEndocrinology
Volume143
Issue number8
DOIs
Publication statusPublished - Aug 2002
Externally publishedYes

Keywords

  • Animals
  • Animals, Newborn
  • Antigens, Neoplasm/analysis
  • Cell Adhesion Molecules/analysis
  • Cell Division
  • Cell Separation/methods
  • Cells, Cultured
  • Epithelial Cell Adhesion Molecule
  • Flow Cytometry
  • Male
  • Rats
  • Rats, Wistar
  • Spermatozoa/physiology

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