A pumpless monolayer microfluidic device based on mesenchymal stem cell-conditioned medium promotes neonatal mouse in vitro spermatogenesis

Student admission to the Atılım University Department of Nutrition and Dietetics started in 2017. Our Department is the only academic institution to offer undergraduate-level education completely in English in the field of Nutrition and Dietetics in Ankara. The studies of our department may be classified into two main categories; education and research. The current education programs are offered taking into consideration the awareness of the responsibility in offering a degree in Nutrition and Dietetics; by competent instructors in the field, and with an inter-disciplinary approach. Our aim for the future alumni of our undergraduate program is to undertake their responsibilities in the light of their information with a professional insight, and the confidence to constantly update themselves at hospitals, polyclinics, public health centers, ministries, catering institutions, food companies, universities and such where they may be employed in positions such as health care professionals, academicians, researchers, directors or policy makers.

Abstract

BackgroundChildhood cancer treatment-induced gonadotoxicity causes permanent infertility/sub-infertility in nearly half of males. The current clinical and experimental approaches are limited to cryopreservation of prepubertal testicular strips and in vitro spermatogenesis which are inadequate to achieve the expanded spermatogonial stem/progenitor cells and spermatogenesis in vitro. Recently, we reported the supportive effect of bone marrow-derived mesenchymal cell co-culture which is inadequate after 14 days of culture in static conditions in prepubertal mouse testis due to lack of microvascular flow and diffusion. Therefore, we generated a novel, pumpless, single polydimethylsiloxane-layered testis-on-chip platform providing a continuous and stabilized microfluidic flow and real-time cellular paracrine contribution of allogeneic bone marrow-derived mesenchymal stem cells.MethodsWe aimed to evaluate the efficacy of this new setup in terms of self-renewal of stem/progenitor cells, spermatogenesis and structural and functional maturation of seminiferous tubules in vitro by measuring the number of undifferentiated and differentiating spermatogonia, spermatocytes, spermatids and tubular growth by histochemical, immunohistochemical, flow cytometric and chromatographic techniques.ResultsBone marrow-derived mesenchymal stem cell-based testis-on-chip platform supported the maintenance of SALL4(+) and PLZF(+) spermatogonial stem/progenitor cells, for 42 days. The new setup improved in vitro spermatogenesis in terms of c-Kit(+) differentiating spermatogonia, VASA(+) total germ cells, the meiotic cells including spermatocytes and spermatids and testicular maturation by increasing testosterone concentration and improved tubular growth for 42 days in comparison with hanging drop and non-mesenchymal stem cell control.ConclusionsFuture fertility preservation for male pediatric cancer survivors depends on the protection/expansion of spermatogonial stem/progenitor cell pool and induction of in vitro spermatogenesis. Our findings demonstrate that a novel bone marrow-derived mesenchymal stem cell-based microfluidic testis-on-chip device supporting the maintenance of stem cells and spermatogenesis in prepubertal mice in vitro. This new, cell therapy-based microfluidic platform may contribute to a safe, precision-based cell and tissue banking protocols for prepubertal fertility restoration in future.

Description

Onen, Selin/0000-0002-3255-3035; KORKUSUZ, PETEK/0000-0002-7553-3915; Kose, Sevil/0000-0003-2188-9534; Gizer, Merve/0000-0003-1911-2363; kulah, haluk/0000-0003-1331-4474