Mesenchymal stem cells differentiation into gamete germ cells by titanium nanotube particles and UV radiation induction

Many of traditional infertility treatment methods due to lack of gamete are ineffective.  Differentiation of stem cells to gamete germ cells may be a good choice for producing gametes for treatment of infertility in clinique. Bone marrow mesenchymal stem cells (BMMSc) were obtained from femoral bone of adult NMRI mice by flushing method. Nano tube particles of titanium (TNT), retinoic acid and UV beam 254 ± 20 nm frequencies were used for differentiation of MSc to gamete germ cells. Ultrastructure of TNT was studied with FEM SE electron microscope. Fibrin coating of Nano tubes was evaluated by SEM. Nature and character of stem cells were measured using cluster– differentiation (CD) by flow cytometry confirmed by CD90, and CD31 markers.  Finally, MVH marker was used for assessment of differentiation of MSc to gamete germ cells with immunohistochemistry assay. Statistical data was analyzed by ANOVA and Mann-Whitney tests. Electron microscopic pictures by FE SEM showed homogenous ultrastructure of titanium nanotubes in 10-15 nm diameters. Coating of TNT by fibrin was confirmed by SEM. MTT assay showed most survival rate of MSc cells in 50 µg/ml concentration of TNT. By CD90 and CD31 markers, nature of the stem cells was controlled by flow cytometry. Finally MVH marker expression showed differentiation of MSc to gamete germ cells in treated as compared to control groups and was confirmed by immunohistochemistry. Application of TNT fibrin coated and UV beam can increase differentiation rate of BMMSc to gamete germ cells significantly as a clinical protocol.