Xenotransplantation of canine spermatogonial stem cells (cSSCs) regulated by FSH promotes spermatogenesis in infertile mice

• Published in: Stem Cell Research & Therapy
• Main Authors: Naira Caroline Godoy Pieri, Ana Carolina Furlanetto Mançanares, Aline Fernanda de Souza, Hugo Fernandes, Angela Maria Gonella Diaza, Fabiana Fernandes Bressan, Kelly Cristine Santos Roballo, Juliana Barbosa Casals, Mario Binelli, Carlos Eduardo Ambrósio, Daniele dos Santos Martins
• Format: Article in Journal/Newspaper
• Language: English
• Published: BMC 2019
• Subjects: Transplant; Spermatogenesis; Hormone; Germ cells; Medicine (General); R5-920; Biochemistry; QD415-436; Canis lupus
• Online Access: https://doi.org/10.1186/s13287-019-1250-9; https://doaj.org/article/1498a524d2204bf490c67c897c6059da

Abstract Background Xenotransplantation of spermatogonial stem cells (SSCs) has become a popular topic in various research fields because manipulating these cells can provide insights into the mechanisms associated with germ cell lines and the entire spermatogenesis process; moreover, these cells can be used in several biotechnology applications. To achieve successful xenotransplantation, the in vitro microenvironment in which SSCs are cultured should be an ideal microenvironment for self-renewal and similar to the in vivo testicular microenvironment. The age of the donor, the correct spermatogenesis cycle, and the quality of the donor tissue are also important. Although cell culture-related factors, such as the in vitro supplementation of hormonal factors, are known to promote successful xenotransplantation in mice, little is known about the influence of these factors on SSCs in vitro or in vivo in other mammalian species, such as dogs (Canis lupus familiaris). In this context, the goals of this study were to test the effect of follicle-stimulating hormone (FSH) on canine spermatogonial stem cell (cSSC) cultures since this hormone is related to the glial cell-derived neurotrophic factor (GDNF) signaling pathway, which is responsible for the self-renewal and maintenance of these cells in vivo, and to investigate the microenvironment of the SSC culture after FSH supplementation. Additionally, in vivo analyses of transplanted FSH-supplemented cSSCs in the testes of infertile mice were performed to assess the capacity of cSSCs to develop, maintain, and restore spermatogenesis. Methods SSCs from canine prepubertal testes (aged 3 months) were cultured in vitro in the presence of FSH (10 IU L−1). GFRA1 transcript expression was detected to confirm the spermatogonia population in culture and the effect of FSH on these cells. The protein and transcript levels of late germ cell markers (GFRA1, DAZL, STRA8, PLZF, and CD49f) and a pluripotency marker (OCT4) were detected at 72 and 120 h to confirm the cSSC phenotype. In ...