Önen, Selin

Prepubertal cancer treatment leads to irreversible infertility in half of the male patients. Current in vitro spermatogenesis protocols and cryopreservation techniques are inadequate to expand spermatogonial stem/progenitor cells (SSPC) from testicles. Bone marrow derived mesenchymal stem cells (BM-MSC) bearing a close resemblance to Sertoli cells, improved spermatogenesis in animal models. We asked if a co-culture setup supported by syngeneic BM-MSC that contributes to the air-liquid interphase (ALI) could lead to survival, expansion and differentiation of SSPCs in vitro. We generated an ALI platform able to provide a real-time cellular paracrine contribution consisting of syngeneic BM-MSCs to neonatal C57BL/6 mice testes. We aimed to evaluate the efficacy of this culture system on SSPC pool expansion and spermatogenesis throughout a complete spermatogenic cycle by measuring the number of total germ cells (GC), the undifferentiated and differentiating spermatogonia, the spermatocytes and the spermatids. Furthermore, we evaluated the testicular cell cycle phases, the tubular and luminal areas using histochemical, immunohistochemical and flow cytometric techniques. Cultures in present of BM-MSCs displayed survival of ID4(+) spermatogonial stem cells (SSC), expansion of SALL4(+) and OCT4(+) SSPCs, VASA(+) total GCs and Ki67(+) proliferative cells at 42 days and an increased number of SCP3(+) spermatocytes and Acrosin(+) spermatids at 28 days. BM-MSCs increased the percentage of mitotic cells within the G2-M phase of the total testicular cell cycle increased for 7 days, preserved the cell viability for 42 days and induced testicular maturation by enlargement of the tubular and luminal area for 42 days in comparison to the control. The percentage of PLZF(+) SSPCs increased within the first 28 days of culture, after which the pool started to get smaller while the number of spermatocytes and spermatids increased simultaneously. Our findings established the efficacy of syngeneic BM-MSCs on the survival and expansion of the SSPC pool and differentiation of spermatogonia to round spermatids during in vitro culture of prepubertal mice testes for 42 days. This method may be helpful in providing alternative cures for male fertility by supporting in vitro differentiated spermatids that can be used for round spermatid injection (ROSI) to female oocyte in animal models. These findings can be further exploited for personalized cellular therapy strategies to cure male infertility of prepubertal cancer survivors in clinics.

This study presents the resonant frequencies of several antennas, which are dual-frequency antennas. These antennas possess two shorting pins to form dual frequency. Among several methods to provide dual operation, two shorting pins are used to form dual frequency equilateral triangular antennas since there are limited studies on this issue and higher frequency ratios are achievable. The present study has been done with this in mind in order to contribute to related literature on such antennas. Here, the parameters that affect the resonant frequencies of the antenna are also evaluated. The thickness, side length, and shorting pin positions are changed numerous times, and the resonant frequencies are examined according to the change of these parameters. Finally, the frequency ratios are determined for each variation, and the results are compared. The proposed antennas in present study achieved a frequency ratio around 5.7, which is significantly higher from the ones presented in the literature.

Journal	Count
Stem Cell Research & Therapy	2
American Journal of Rhinology & Allergy	1
Journal of Assisted Reproduction and Genetics	1
Journal of Hazardous Materials	1
Progress In Electromagnetics Research Symposium (PIERS) -- AUG 19-23, 2012 -- Moscow, RUSSIA	1

Önen, Selin

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