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Differentiation of Murine Premigratory Primordial Germ Cells in Culture¹

^a Department of Molecular Genetics and Microbiology, University of Florida College of Medicine, Gainesville, Florida 32610-0266 ^b Department of Anatomy and Cell Biology, School of Medicine, University of Kansas Medical Center, Kansas City, Kansas 66160-7400

In the mouse embryo, primordial germ cells first appear in the extraembryonic mesoderm and divide rapidly while migrating to the fetal gonad. Shortly after their arrival in the gonad, germ cells sexually differentiate as proliferation ceases. Previous studies have established that primordial germ cells proliferate and migrate in feeder layer culture. To explore cellular regulation of fetal germ cell development, we have used germ cell nuclear antigen 1 (GCNA1), a marker normally expressed only in postmigratory germ cells, to investigate the developmental potency of both pre- and postmigratory cells in this culture system. We found that explanted premigratory germ cells will initiate expression of this marker and are, therefore, capable of undertaking some aspects of gonocyte differentiation without intimate exposure to the fetal gonad. We have also tested whether postmigratory gonocytes are stable in culture. As detected by either alkaline phosphatase or GCNA1, we did not detect long-term survival of either prospermatogonia or oogonia under conditions that support the survival, proliferation, and differentiation of earlier premigratory cells. These observations are consistent with an autonomous cellular mechanism governing the initial stages of gonocyte differentiation, and suggest that differentiation towards gonocytes is accompanied by a change in requirements for cell survival.

¹ This work was supported by a New Investigator award from the Cooperative States Research Service, U.S. Department of Agriculture (93–37205–9074), and by a College Incentive Fund award from the University of Florida College of Medicine to J.R.

² Correspondence: James Resnick, Department of Molecular Genetics and Microbiology, University of Florida, 1600 SW Archer Road, Gainesville, FL 32610–0266. FAX: 352 392 3133; resnick{at}college.med.ufl.edu

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