Rat Sertoli cells express epithelial but also mesenchymal genes after immortalization with SV40.
A new immortal Sertoli cell line from pubertal rat testis was established and characterized. We have generated the clonal line SCIT-C8 expressing established markers for Sertoli cells (SC) like transferrin, clusterin and steel factor/stem cell factor (SCF). Additionally, the immortalized cells express afadin, a protein which is a member of tight and adherens junctions, therefore the cells may be useful for studies of the blood-testis barrier (BTB) in vitro. In contrast to primary SC, the immortalized cells lost expression of androgen receptor and responsiveness to androgens and follicle-stimulating hormone. Surprisingly, we found mRNA expression and protein secretion of the mesenchymal markers, fibronectin and entactin-1, which we also observed for the immortalized SC lines, ASC-17D and 93RS2. In comparison to primary SC, the immortalized cells demonstrated enhanced adhesion in vitro. This correlated with the expression of entactin-1 because adhesion was strongly reduced by antibody perturbation experiments. Additionally, we found the alternatively spliced and primarily muscle cell-specific long variant of TGF-beta2 not only in peritubular cells (PC), but also in the primary and immortalized SC. Furthermore, all immortalized cell lines secreted higher amounts of TGF-beta2 than primary SC. In conclusion, the immortalized SC lines from different developmental stages showed a similar pattern of epithelial and mesenchymal markers.
Konrad
Lutz
L
Department of Urology, Uniklinikum Lahnberge, Baldingerstr, D-35033 Marburg, Germany. Konrad@post.med.uni-marburg.de
Munir Keilani
Marcel
M
Cordes
Andrea
A
Völck-Badouin
Elke
E
Laible
Leslie
L
Albrecht
Martin
M
Renneberg
Heiner
H
Aumüller
Gerhard
G
D000818:Animals; D000952:Antigens, Polyomavirus Transforming / Q000235:genetics / Q000378:metabolism; D002461:Cell Line, Transformed; D004848:Epithelium / Q000502:physiology*; D005109:Extracellular Matrix / Q000737:chemistry; D005353:Fibronectins / Q000235:genetics / Q000378:metabolism; D008297:Male; D008562:Membrane Glycoproteins / Q000235:genetics / Q000378:metabolism; D008648:Mesoderm / Q000502:physiology*; D051381:Rats; D012708:Sertoli Cells / Q000166:cytology* / Q000502:physiology*; D013737:Testis / Q000166:cytology
D000952:Antigens, Polyomavirus Transforming; D005353:Fibronectins; D008562:Membrane Glycoproteins; C029967:nidogen