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A three-dimensional organoid culture model to study SSC maintenance and spermatogenesis in vitro



Dr Kevin Coward


Annually, around 2000 new cases of childhood cancer are diagnosed in the UK. Prepubertal boys are often rendered infertile by aggressive forms of chemo- or radiotherapy. The production of functional sperm from spermatogonial stem cells (SSCs) in the testis is only initiated at puberty. Consequently, the only way to preserve fertility in these patients is to cryopreserve fragments of testicular tissue for subsequent culture in vitro. Within the testes, SSCs are located inside a specialized three-dimensional (3D) structure (the seminiferous tubules); direct cell-to-cell interactions play a critical role in their proliferation and development.

In recent decades, the extracellular matrix (ECM) has been widely used in cell culture to provide a 3D scaffold-based environment to mimic the in vivo conditions. When designing 3D micro-tissues, it is important to recapitulate the microenvironment experienced in vivo. This can be achieved by co-culturing multiple cell types within artificial tissue matrices or scaffolds. Organoids are self-organizing multicellular spheroids that form in 3D culture systems and are similar to actual organs. The capacity of organoids to generate complex 3D structures resembling organs is revolutionizing the fields of developmental and stem cell biology. 

Using Matrigel as an ECM, structural testis organoids have been found to secrete high levels of testosterone and inhibin B over 12 weeks with preserved responsivity to gonadotropins. Cellular self-assembly and organoid formation were achieved independent of the culture microenvironment, thus providing a template for studying testicular organoid self-assembly and endocrine function and a platform for improving the engineering of functional testicular tissues. The organoids generated from immature testicular cells consist of SSCs and have the potential to develop into mature sperm. However the specific modes of regulation inside the SSC niches have not yet been elucidated. Furthermore, whether multipotency and organoid formation is restricted to a certain type of cells is unknown. This project aims to enhance previous work carried out in Dr Coward’s laboratory towards the development of a 3D scaffolding organoid system to support the proliferation and differentiation of SSCs.


Cell culture including 2D & 3D culture, immunofluorescence and immunocytochemistry, confocal microscopy, RT-qPCR , Flow cytometry.

As well as the specific training detailed above, students will have access to a wide-range of seminars and training opportunities through the many research institutes and centres based in Oxford.