An in vitro study of the impact of 4mT static magnetic field to modify the differentiation rate of rat bone marrow stem cells into primordial germ cells Fatemeh Javani Jouni a , Parviz Abdolmaleki a,n , Mehrdad Behmanesh b , Mansoureh Movahedin c a Department of Biophysics, Faculty of Biological Sciences, Tarbiat Modares University (TMU), P.O.B 14115-154, Tehran, Iran b Department of Genetics, Faculty of Biological Sciences. Tarbiat Modares University (TMU), P.O.B 14115-154, Tehran, Iran c Department of Anatomical sciences, Medical Sciences Faculty, Tarbiat Modares University (TMU), P.O.B 14115-175, Tehran, Iran article info Article history: Received 22 January 2014 Received in revised form 13 June 2014 Accepted 18 June 2014 Keywords: Bone marrow stem cell Bone morphogenetic protein 4 Differentiation Primordial germ cell Static magnetic field abstract This investigation was performed to evaluate the differentiation capacity and alteration in genes expression patterns during in vitro differentiation of bone marrow stem cells into primordial germ cells using static magnetic field (4 mT) and BMP-4 (25 ng/ml). The rate of differentiation was investigated using the Real Time-PCR method for tracing expression of differentiation markers (Oct-4, Nanog, C-Myc, Fragilis, Mvh and Stella). Then, immunocytochemical reaction was carried out for detection of marker proteins (Oct4 and Mvh). Increasing of the exposure time of the 4 mT SMF (24 and 48 h) and treatment time with 25 ng/ml BMP4 (48 and 96 h) indicated a marked decrease in expression of pluripotency genes (Oct-4, Nanog and C-Myc) and Oct4 protein and increase in primordial germ cell-specific genes (Fragilis, Mvh and Stella) and Mvh protein compared with the control group. Final results showed that in a synergistic manner, the combination of SMF with BMP4 exaggerates the differentiation potential of BMSCs to PGCs by activating the MAPK pathway and altering the Ca 2 þ concentration. & 2014 International Society of Differentiation. Published by Elsevier B.V. All rights reserved. 1. Introduction Natural magnetic fields (MFs) vary over the earth's surface based on the geographic longitude and latitude of the locations between 0.035 and 0.07 mT. This natural MF is sensed by certain animals that use it for orientation (Bekhite et al., 2013). The interest in the variation biological effects of non-ionizing electromagnetic fields (EMFs) on the whole organism, as well as on cellular systems, has considerably increased in recent years in consideration of their probable health risk for humans (Pagliara et al., 2009). Several studies have shown that MF has an influence on a large variety of cellular functions; nevertheless the its exact mechanism(s) still is not clear (Marędziak et al., 2014). Few studies exist on the effects of static magnetic fields (SMFs) on living cells and tissues, compared with low-frequency magnetic fields. Most of these studies dealt with the potentially genotoxic or oncogenic effects especially those that have a relationship with clinical applications of static magnetic field. Part of these studies focused on this basic idea that SMF play a critical role in activating and/or alternating the molecular mechan- isms in eukaryotic cells (Miyakoshi, 2005). Many experiments were recently performed to study the interaction between organisms and MFs (Fanelli et al., 1999; Lacy-Hulbert et al., 1998). A recent article demonstrated that cardiomyogenesis of Flk-1 þ cardiac progenitor cells derived from mouse embryonic stem cells (ESs) can be enhanced by application of static MFs (0.3 to 5 mT). Moreover, the relationship between MF-mediated intracellular ROS generation and [Ca 2 þ ] as second messengers in signal pathways leading to cardiomyocytes differentiation is not clear (Bekhite et al., 2013). Another study investigated the effects of MFs of 50 Hz and 1.1 mT on the differentiation and intracellular free calcium of bone marrow mesenchymal stem cells (Zhao et al., 2008). They showed that MFs enhanced the cellular differentiation and the intracellular free Ca 2 þ concentration in MSCs. Converging data indicate that the primary site of action of magnetic fields is the plasma membrane (Rosen, 2003a, 2003b). The influence exerted by MFs (static or oscillating) on the plasma membrane has been described at different levels on: the plasma membrane surface (Paradisi et al., 1993), the distribution of Contents lists available at ScienceDirect journal homepage: www.elsevier.com/locate/diff Differentiation http://dx.doi.org/10.1016/j.diff.2014.06.001 Join the International Society for Differentiation (www.isdifferentiation.org) 0301-4681/& 2014 International Society of Differentiation. Published by Elsevier B.V. All rights reserved. n Corresponding author. Tel.: þ98 21 82883404; fax: þ98 21 82884717. E-mail addresses: Javani@modares.ac.ir (F.J. Jouni), Parviz@modares.ac.ir (P. Abdolmaleki), Behmanesh@modares.ac.ir (M. Behmanesh), Movahed.m@modares.ac.ir (M. Movahedin). Please cite this article as: Jouni, F.J., et al., An in vitro study of the impact of 4mT static magnetic field to modify the differentiation rate of rat bone marrow stem cells into primordial germ cells. Differentiation (2014), http://dx.doi.org/10.1016/j.diff.2014.06.001i Differentiation ∎ (∎∎∎∎) ∎∎∎–∎∎∎