Fig. 3
BMSC-contributed MFD provides maintenance and enlargement of SALL4(+) and PLZF(+) SSPC pool and PAS-stained spermatogonia in vitro. Immune labeling of a SALL4(+) and c PLZF(+) SSPCs, and e PAS-stained of spermatogonia in prepubertal mice testes from day 7 to 42 in BMSC-CM-applied and non-applied HD and MFD (SALL4 IHC and PAS–hematoxylin, 1000x). Note the presence of SSPCs indicated by arrow on SALL4-labeled sections. In time change in b number of SALL4(+) cells, d ratio of PLZF(+) cells to total testicular cells and f spermatogonium count in BMSC-CM-applied and non-applied groups are illustrated in bar graph with standard deviation and data distribution (*p < 0.05, n = 6 testes, 50 tubules for SALL4; n = 6 testes for PLZF). g Line graph illustrates positive correlation of SALL4 with PLZF labeling and spermatogonium number for SSPCs in control and BMSC groups (R2 = 0.0319, p = 0.5785; R2 = 0.9994, p = 0.001, respectively) (arrows: SSPCs, asterisks: seminiferous tubules)