Fig. 4
Effect of CBD and/or THC on proliferation, apoptosis, and germ cell numbers. Density of (A) proliferating cells (KI67 +) and (B) apoptotic cells (Cas3 +) in (A,B) extracordonal and (C,D) intracordonal tissue after CBD and/or THC treatment at 10−5 M. Results of cell quantification normalized by surface unit and expressed as fold from respective controls in 8–10 and 10–12 DW embryonic testes. Data presented as means + / − SEM. An ANOVA test was performed followed by a Wilcoxon test for pair comparisons (*, p < 0.05; **, p < 0.01). (E) Representative immunostaining of LIN28, a specific marker of germ cell in control (DMSO/EtOH) and CBD/THC at 10−5 M treated explants from a 10–12 DW human fetal testis. LIN28 appears dark brown (DAB staining) in the cytoplasm of germ cells within the intracordonal tissue. Sections were counterstained with hematoxylin (in blue). No modification of germ cell morphology was observed. (F) Density of LIN28 positive cells (LIN28 +) in intracordonal tissue after CBD and/or THC treatment at 10−5 M compared to respective controls in 8–10 and 10–12 DW embryonic testis. Scale bar = 50 µm