A Role for Exchange of Extracellular Vesicles in Porcine Spermatogonial Co-Culture
Spermatogonial stem cells (SSCs) provide the basis for lifelong male fertility through self-renewal and differentiation. Prepubertal male cancer patients may be rendered infertile by gonadotoxic chemotherapy and, unlike sexually mature men, cannot store sperm. Alternatively, testicular biopsies take...
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| Format: | Article |
| Language: | English |
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MDPI AG
2022-04-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/23/9/4535 |
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| author | Shiama Thiageswaran Heather Steele Anna Laura Voigt Ina Dobrinski |
| author_facet | Shiama Thiageswaran Heather Steele Anna Laura Voigt Ina Dobrinski |
| author_sort | Shiama Thiageswaran |
| collection | DOAJ |
| description | Spermatogonial stem cells (SSCs) provide the basis for lifelong male fertility through self-renewal and differentiation. Prepubertal male cancer patients may be rendered infertile by gonadotoxic chemotherapy and, unlike sexually mature men, cannot store sperm. Alternatively, testicular biopsies taken prior to treatment may be used to restore fertility in adulthood. Testicular SSC populations are limited, and in vitro culture systems are required to increase numbers of SSCs for treatment, demanding culture systems for SSC propagation. Using the pig as a non-rodent model, we developed culture systems to expand spermatogonia from immature testis tissue, comparing different feeders (Sertoli cells, peritubular myoid cells (PMCs) and pig fetal fibroblasts (PFFs)). Spermatogonia co-cultured with Sertoli cells, PMCs and PFFs had comparable rates of proliferation and apoptosis. To elucidate the mechanism behind the beneficial nature of feeder layers, we investigated the role of extracellular vesicles in crosstalk between spermatogonia and feeder cells. Sertoli cell-released exosomes are incorporated by spermatogonia, and inhibition of exosomal release reduces spermatogonial proliferation. Together, these results show that PMCs, PFFs and Sertoli cells promote spermatogonial proliferation in co-culture, with exosomal exchange representing one possible mechanism. Further characterization of exosomal cargo may ultimately allow the development of feeder-free culture systems for clinical use. |
| first_indexed | 2024-03-10T04:08:44Z |
| format | Article |
| id | doaj.art-f5ed3ae4b3874ed29656f7ce31da47dc |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-10T04:08:44Z |
| publishDate | 2022-04-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-f5ed3ae4b3874ed29656f7ce31da47dc2023-11-23T08:18:37ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672022-04-01239453510.3390/ijms23094535A Role for Exchange of Extracellular Vesicles in Porcine Spermatogonial Co-CultureShiama Thiageswaran0Heather Steele1Anna Laura Voigt2Ina Dobrinski3Department of Biochemistry and Molecular Biology, Cumming School of Medicine, University of Calgary, Calgary, AB T2N 4N1, CanadaDepartment of Comparative Biology and Experimental Medicine, Faculty of Veterinary Medicine, University of Calgary, Calgary, AB T2N 1N4, CanadaDepartment of Comparative Biology and Experimental Medicine, Faculty of Veterinary Medicine, University of Calgary, Calgary, AB T2N 1N4, CanadaDepartment of Biochemistry and Molecular Biology, Cumming School of Medicine, University of Calgary, Calgary, AB T2N 4N1, CanadaSpermatogonial stem cells (SSCs) provide the basis for lifelong male fertility through self-renewal and differentiation. Prepubertal male cancer patients may be rendered infertile by gonadotoxic chemotherapy and, unlike sexually mature men, cannot store sperm. Alternatively, testicular biopsies taken prior to treatment may be used to restore fertility in adulthood. Testicular SSC populations are limited, and in vitro culture systems are required to increase numbers of SSCs for treatment, demanding culture systems for SSC propagation. Using the pig as a non-rodent model, we developed culture systems to expand spermatogonia from immature testis tissue, comparing different feeders (Sertoli cells, peritubular myoid cells (PMCs) and pig fetal fibroblasts (PFFs)). Spermatogonia co-cultured with Sertoli cells, PMCs and PFFs had comparable rates of proliferation and apoptosis. To elucidate the mechanism behind the beneficial nature of feeder layers, we investigated the role of extracellular vesicles in crosstalk between spermatogonia and feeder cells. Sertoli cell-released exosomes are incorporated by spermatogonia, and inhibition of exosomal release reduces spermatogonial proliferation. Together, these results show that PMCs, PFFs and Sertoli cells promote spermatogonial proliferation in co-culture, with exosomal exchange representing one possible mechanism. Further characterization of exosomal cargo may ultimately allow the development of feeder-free culture systems for clinical use.https://www.mdpi.com/1422-0067/23/9/4535spermatogonial stem cellsspermatogoniaco-culturesextracellular vesiclesexosomespig |
| spellingShingle | Shiama Thiageswaran Heather Steele Anna Laura Voigt Ina Dobrinski A Role for Exchange of Extracellular Vesicles in Porcine Spermatogonial Co-Culture International Journal of Molecular Sciences spermatogonial stem cells spermatogonia co-cultures extracellular vesicles exosomes pig |
| title | A Role for Exchange of Extracellular Vesicles in Porcine Spermatogonial Co-Culture |
| title_full | A Role for Exchange of Extracellular Vesicles in Porcine Spermatogonial Co-Culture |
| title_fullStr | A Role for Exchange of Extracellular Vesicles in Porcine Spermatogonial Co-Culture |
| title_full_unstemmed | A Role for Exchange of Extracellular Vesicles in Porcine Spermatogonial Co-Culture |
| title_short | A Role for Exchange of Extracellular Vesicles in Porcine Spermatogonial Co-Culture |
| title_sort | role for exchange of extracellular vesicles in porcine spermatogonial co culture |
| topic | spermatogonial stem cells spermatogonia co-cultures extracellular vesicles exosomes pig |
| url | https://www.mdpi.com/1422-0067/23/9/4535 |
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