following subcutaneous autograft (12.01AE0.87ng/DL (neg CTL) vs 15.1AE1.50ng/DL (15 days Autograft) vs 22.26AE1.33ng/DL (30 days Autograft) vs 37.3AE9.6ng/DL (60 days Autograft)), demonstrating differentiating LSC within the autograft. In addition, we found that levels of 3BHSD were induced upon SAG (DHH inducer) treatment in-vitro conditions. Immunostaining autografts (4 weeks), containing LSCs treated with SAG before subcutaneous implantation, showed higher levels of SOX9, and a-SMA establishing that Hedgehog signalling induces survival of adjacent testicular cells that are required for graft survival and LSC differentiation.CONCLUSIONS: Our results demonstrate that subcutaneous autograft of LSC in combination with Sertoli cells and myoid cells can increase serum testosterone without inhibiting circulating LH and FSH in castrate mice. Extratesticular LSC appear to be regulated by Hedgehog signalling. Leydig stem cell autograft can a novel therapeutic approach to increase serum testosterone while simultaneously preserving hypothalamic-pituitary-gonadal axis. Factors involved in hedgehog signalling can be utilized to enhance graft efficacy.
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