The effects of melatonin on colonization of neonate spermatogonial mouse stem cells in a three-dimensional soft agar culture system

Abstract

BackgroundMelatonin is a pleiotropic hormone with powerful antioxidant activity both in vivo and in vitro. The present study aimed to investigate the effects of melatonin on the proliferation efficiency of neonatal mouse spermatogonial stem cells (SSCs) using a three-dimensional soft agar culture system (SACS) which has the capacity to induce development of SSCs similar to in vivo conditions.MethodsSSCs were isolated from testes of neonate mice and their purities were assessed by flow cytometry using PLZF antibody. Isolated testicular cells were cultured in the upper layer of the SACS in αMEM medium in the absence or presence of melatonin extract for 4 weeks.ResultsThe identity of colonies was confirmed by alkaline phosphatase staining and immunocytochemistry using PLZF and α6 integrin antibodies. The number and diameter of colonies of SSCs in the upper layer were evaluated at days 14 and 28 of culture. The number and diameter of colonies of SSCs were significantly higher in the melatonin group compared with the control group. The levels of expression of ID-4 and Plzf, unlike c-kit, were significantly higher in the melatonin group than in the control group.ConclusionsResults of the present study show that supplementation of the culture medium (SACS) with 100 μM melatonin significantly decreased reactive oxygen species (ROS) production in the treated group compared with the control group, and increased SSC proliferation.

Proliferation and Differentiation of Mouse Spermatogonial Stem Cells on a Three-Dimensional Surface Composed of PCL/Gel Nanofibers

“…Due to paracrine secretion and supportive role of Sertoli cells in the survival, proliferation and fate determination of SSCs (Griswold, 1998), we presume that Sertoli cells grown on the nanofibrous scaffold were more functional and created a more appropriate microenvironment for SSCs proliferation and colonization. In agreement with our findings, many studies have reported increased number and diameter of colonies of SSC by improving the quality of culture condition (Baazm et al, 2013;Aliakbari et al, 2017;Navid et al).…”

“…Due to presence of SSCs on the basement membrane in a 3D microenvironment, the extracellular matrix may play an important role in the behavior of these cells. Hence, various 3D culture systems such as soft agar culture system (SACS), methylcellulose culture system (MCS), collagen gel matrix and alginate capsule have been developed for in vitro proliferation and differentiation of SSCs (Lee et al, 2006;Huleihel et al, 2015;Jalayeri et al, 2017;Navid et al, 2017).…”

Differentiation of spermatogonial stem cells by soft agar three-dimensional culture system

“…In contrast to our study, the study of Eslahi et al (2013) showed that the number of mouse SSCs colonies significantly decreased in the first and third weeks of culture when cultured on poly-L-lactic acid (PLLA) nanofiber in comparison with the control group [30]. In the study of Navid et al (2017), the number of mouse spermatogonial colonies significantly increased in SACS culture system compared to two-dimensional culture system. Increasing the number of colonies in our study is in agreement with the study of Navid et al [31].…”

The effects of human menstrual blood stem cells‐derived granulosa cells on ovarian follicle formation in a rat model of premature ovarian failure

“…Primers were designed by the allele ID software. The sequences of primers are shown in Table (Navid, Abbasi, & Hoshino, ).…”

“…The expression of specific ovarian granulosa cell genes such as Amh The sequences of primers are shown in Table 1 (Navid, Abbasi, & Hoshino, 2017).…”