Figure 4. Assessment of cellular proliferation after PCNA (Proliferating Cell Nuclear Antigen) and Tra98 immunodetection on serial sections of seminiferous tubules in organotypic culture of fresh and frozen-thawed pre-pubertal mouse testicular tissue.

(A) Cellular proliferation assessment for organotypic culture of fresh immature mice testicular tissue assessed after 0 (1 and 4) or 11 days (2,3,5,6) of culture with RE6 (2 and 5) or RERA5 (3 and 6)

(B) Cellular proliferation assessment for organotypic culture of frozen-thawed testicular tissue with stabilisation phase at –8°C (2 and 5) and -9°C (3 and 6) compared to fresh control organotypic culture (1 and 4)

1 to 3 : PCNA immunodetection

4 to 6 : Tra98 iommunostaining

** : Germ cells PCNA positive ; ¤¤ : Sertoli cell PCNA positive ; ^^ : Sertoli cell PCNA negative

Footnotes:

RE6 : 10^-6M retinol ; RERA5 : 10^-5M retinoic acid and 3.3.10^-7M retinol