Reconstructed seminiferous tubules from ectopic grafting of testicular cells / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the ectopic grafts of mouse testicular cells by observing the reconstruction of seminiferous tubules, colonization of spermatogenic cells and spermatogenesis using immunodeficient mice as recipients.</p><p><b>METHODS</b>The testes of newborn male ICR mice were digested to obtain single cell suspension. The cells were then mixed with matrigel and subcutaneously grafted into the dorsal region of the male nude mice. The mice were castrated after the operation and the grafts were dissected from 5 of the nude mice at 4, 6, 8 and 10 weeks, respectively. The success rates of transplantation and the graft diameters were calculated, and the structure of the reconstituted seminiferous tubules, colonization of the germ cells and spermatogenesis were observed by HE staining and immunohistochemistry.</p><p><b>RESULTS</b>All the mice recipients survived after the testicular cell transplantation. Within 10 weeks after the operation, tissue masses could be observed, with the diameter increased from (3.91 +/- 0.71) mm at 4 weeks to (6.69 +/- 0.50) mm. Neovascularization was detected at the surface of the masses and seminiferous tubule structures found in the grafts. The germ cells that developed from spermatogonia to round spermatids were observed, but with no sperm in the tubules. Germ cells, Sertoli cells and Leydig cells were identified by immunochemical detection of Mvh, Gata4 and P450Scc in the grafts at 8 weeks.</p><p><b>CONCLUSION</b>Seminiferous tubules could be ectopically reconstructed from suspension of neonatal mouse testicular cells. Ectopic grafting provided a preferable model for the studies on testis tissue engineering and interactions between testicular cells during testicular development and spermatogenesis.</p>

Spontaneous differentiation potency of induced pluripotent stem cells into male germ cells in vitro / 中华男科学杂志

<p><b>OBJECTIVE</b>To explore the expression profile of male germ cell-associated genes during the spontaneous differentiation of induced pluripotent stem cells (iPS) and assess the potency of their spontaneous differentiation into male germ cells in vitro.</p><p><b>METHODS</b>Embryoid body (EB) formation was used to promote the spontaneous differentiation of iPS into male germ cells, and the expressions of germ cell-associated genes were detected by real-time PCR and PCR.</p><p><b>RESULTS</b>Real-time PCR and PCR revealed different expression levels of relevant genes at different times of iPS spontaneous differentiation into male germ cells. Each of the 9 genes analyzed exhibited one of the four temporal expression patterns: wavelike increase of Oct4, progressive decrease of Dppa3 and Stra8, wavelike decrease of Dazl, and decrease following initial increase of Tex14, Msy2, Scp1, Scp3 and Akap3.</p><p><b>CONCLUSION</b>Induced pluripotent stem cells express male germ cell-associated genes and male haploid genes during their spontaneous differentiation through EB formation, and have the potency of differentiating into male gametes.</p>