Allojenik Mezenkimal Kök Hücreler ve Mikroakışkan Sistemle Vücut Dışında Testisteki Spermatogonyal Kök Hücre Mikroçevresinin Devamlılığının Sağlanması

Abstract

Chemoradiotherapeutics used in childhood cancers result in permenant infertility in half of male patients. Bone marrow derived mesenchymal stem cells (BMMSC) share some of their secretomes and their embryonic origin with Sertoli cells which are main modulators of spermatogenesis. Microfluidic devices (MFD) are technologic platforms that simulate microvascular flow in the body. The study has been conducted as a randomized, prospective, experimental, interdisciplinary research. Dependent variables are the measurement parameters; the independent variables are the groups (MFD and hanging drop (HD); usage and not usage of MSC secretions) and the time (7, 28 and 42 days). Tubular epithelial thickness, luminal diameter, number of spermatogonium, spermatocyte and spermatid were higher in BMMSC-CM and MFD groups when compared to control (p<0.05). SALL4 immune labeled SSCs increased with use of BMMSC-CM and MFD (p<0.05). These results are confirmed by flow cytometric analyses of PLZF (SSCs), c-Kit (spermatogenic cell series) and VASA (germ cells) labeled cells (p<0.005). Testosterone level is increased with use of BMMSC-CM and MFD on days 7 and 42 by mass spectrometry comparing to control (p<0.005). As a result, through the BMMSC-CM supported, novel, technological MFD, seminiferous tubular epithelial thickness, luminal diameter, testosterone level, SSC and spermatogenic cell pool have been increased during 42-day long in vitro culture period. With the data obtained from this study, the supportive effect of BMMSC-CM contributed MFD in in vitro spermatogenesis is demonstrated. This new technology has the potential of being a new therapeutic device for ensuring the continuity of fertility in male prepubertal cancer patients. This model may be a solution for decreasing the economic dependency of our nation to the foreign countries by solving an important health problem like male infertility.