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Fig. 5 | Stem Cell Research & Therapy

Fig. 5

From: Previously claimed male germline stem cells from porcine testis are actually progenitor Leydig cells

Fig. 5

Identification of the C2 clusters as progenitor Leydig cells (PLCs) derived from the porcine testis. a RT-PCR analysis showed that the C2 expressed Leydig cells (LC)-specific genes, such as Gata4, Pdgfrα1, Lifr, Cyp11α1, Cyp17α1, and Star, but did not express the Sertoli cell (SC)-specific marker Sox9 or the peritubular myoid cell (PTM) marker α-Sma. b Images show that, by day 7 of culture without germ cells, the testicular fibroblasts formed clusters that appeared similar to the C2. c Immunofluorescent staining showed that GATA4, PDGFRA1, LIFR, NESTIN, CYP11A1, CYP17A1, and StAR, were all expressed in the C2. The nuclei were stained with Hoechst 33,342 (blue). d Immunohistochemistry staining showed the expression levels of PDGFRA1, LIFR, NESTIN, CYP11A1, CYP17A1, StAR, and 3β-HSD in porcine testicular tissues from 5-day-old piglets. e Flow cytometric analyses showed the expression of mesenchymal stem cell-specific markers (CD29, CD44, CD51, CD73, and CD105) but no expression of hematopoietic marker CD45 in the C2. f Multipotency analyses showed that the C2 maintained multipotency after several passages and displayed strong AP activity. Immunofluorescent staining showed that the C2 expressed PDGFRA1 (Red), but not 3β-HSD. The nuclei were stained with Hoechst 33,342 (blue). Oil red and Alizarin red staining showed they had the capacity to differentiate into adipogenic and osteogenic lineages, respectively. g Images of the C2 following adult Leydig cell (ALC) differentiation induction. h Immunofluorescent staining showed 3β-HSD expression in the induced ALCs. i The testosterone concentration was increased during the induction of ALC differentiation (n = 6). The data are expressed as mean ± SEM

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