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

Stem Cell Research & Therapy

Research

Haluk Külah¹  Ali Can Atik¹  Petek Korkusuz²  Sevil Köse³  Merve Gizer⁴  Selin Önen⁵  Önder Yaman⁶ 

[1] Department of Electrical and Electronics Engineering, Middle East Technical University, Ankara, Turkey;METU MEMS Center, Ankara, Turkey;Department of Histology and Embryology, Faculty of Medicine, Hacettepe University, 06100, Sihhiye, Ankara, Turkey;Department of Plastic, Reconstructive, and Aesthetic Surgery, Akdeniz University, Antalya, Turkey;Department of Stem Cell Sciences, Hacettepe University, Ankara, Turkey;Department of Stem Cell Sciences, Hacettepe University, Ankara, Turkey;Department of Medical Biology, Atilim University, Ankara, Turkey;Department of Urology, Ankara University, Ankara, Turkey;

关键词: In vitro spermatogenesis;    Microfluidics;    Mesenchymal stem cells;    Male infertility;    Prepubertal boys;   

DOI  :  10.1186/s13287-023-03356-x

 received in 2023-03-06, accepted in 2023-04-27,  发布年份 2023

来源: Springer

【 摘 要 】

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.

【 授权许可】

CC BY   
© The Author(s) 2023

【 图 表 】

41116_2023_36_Article_IEq447.gif

41116_2023_36_Article_IEq437.gif

41116_2023_36_Article_IEq425.gif

41116_2023_36_Article_IEq392.gif

41116_2023_36_Article_IEq366.gif

Fig. 3