Fig. 4

Restoration of fertility in T-BST-75 mice after GCT. a Hematoxylin and eosin-stained testicular sections of T-BST-75-GCT mice. Complete stages of spermatogenesis were observed in many tubules. Several tubules only showed initial stages of spermatogenesis. Few tubules were also seen in which spermatogenesis was not started. Black arrowhead marks the sperm, green arrowhead marks the proliferating Gc. Scale bar: 100 μm. b A magnified image of hematoxylin and eosin-stained testicular sections of T-BST-75-GCT mice. Black arrowhead shows two Gc which were harbored in a single evacuated space, yellow arrowhead marks the basement membrane. Scale bar: 100 μm. c Mean testis weight of T-BST-75-GCT mice as compared to non-transplanted control mice. A significant (p < 0.0001) increase in testes weight was observed in the transplanted testes (n = 10). d Mean sperm count of T-BST-75-GCT mice as compared to non-transplanted control mice. A significant (p < 0.0001) increase in sperm count was observed in the transplanted testes (n = 10). e In vivo GFP expression in T-BST-75-GCT testes as compared to non-transplanted control testes 2 months after GCT. GFP expression was observed in transplanted testes. i Image under bright field stereo-zoom microscope. ii Image under UV with FITC filter. Yellow dotted line marks the transplanted testis. White dotted line marks the non-transplanted testis. f Western blot of total protein isolated from T-BST-75-GCT testes as compared to wild-type testis. Transplanted testis showed presence of 28 KDa protein corresponding to GFP. β-actin was used as internal control. g Immunostaining showing GFP expression in the Gc of seminiferous tubules of T-BST-75-GCT testis. GFP expression was observed specifically in Gc of the testis. i and ii shows testicular sections of GCT testis. iii and iv shows testicular sections of non-transplanted testes. Yellow arrowhead shows GFP expression in the Gc. Scale bar: 100 μm. All data were recorded 2 months after GCT