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Establishment of Cell Lines with Rat Spermatogonial Stem Cell Characteristics

Department of Endocrinology (A.M.M.v.P., H.L.R.-G., L.B.C., D.G.d.R.), University of Utrecht, 3584 CH Utrecht, The Netherlands; Department of Cell Biology, University Medical Center Utrecht, 3584 CX Utrecht, The Netherlands; Department of Radiotherapy (I.S.G.), University Medical Center Utrecht, 3584 CX Utrecht, The Netherlands; Department of Biochemistry (F.M.F.v.D.-E.), Cell Biology and Histology, Veterinary School, University of Utrecht, 3584 CL Utrecht, The Netherlands

Address all correspondence and requests for reprints to: Dr. A. M. M. van Pelt, Department of Cell Biology, University Medical Center, location AZU, HP G02.525, Heidelberglaan 100, 3584 CX Utrecht, The Netherlands. E-mail: . a.m.m.vanpelt{at}med.uu.nl

Spermatogonial cell lines were established by transfecting a mixed population of purified rat A_s (stem cells), A_pr and A_al spermatogonia with SV40 large T antigen. Two cell lines were characterized and found to express Hsp90 and oct-4, specific markers for germ cells and A spermatogonia, respectively. Expression of c-kit, normally expressed in A spermatogonia from late A_al spermatogonia onwards, could not be detected in either cell line. Furthermore, no expression of vimentin (Sertoli cell marker) and -smooth muscle actin (peritubular cell marker) could be found. Upon transplantation of these cell lines into recipient mice, the cells were found to be able to migrate to the basement membrane and to colonize seminiferous tubules. Taken together, we conclude that our cell lines have spermatogonial stem cell characteristics. These first spermatogonial cell lines with stem cell characteristics can now be used to study spermatogonial gene expression in comparison with more advanced germ cells.

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