Promoting mBMMSC differentiation into late-stage germ-like cells through retinoic acid, fibrin-coated titanium nanotubes, and ultraviolet radiation.

The primary objective of advancements in stem cell biology for reproductive medicine has been the production of artificial gametes from multipotent stem cells. We examined the efficacy of upregulating germline stem cell markers to convert mouse bone marrow mesenchymal stem cells (mBMMSCs) into late-stage germ-like cells (GLCs). The multipotent differentiation potential of mBMMSCs was investigated using oil red-O and alizarin red-S staining. The differentiation of mBMMSCs into GLCs was also investigated in relation to the effects of high concentrations of retinoic acid, ultraviolet (UV) light, and titanium nanotubes (TNTs) coated with fibrin (F). The biocompatibility and morphology of TNT, as well as the characteristics of F+TNT, were investigated through the use of MTT and scanning electron microscopy (SEM) experiments. After 14 days, the optimal TNT concentration for differentiation was 50 µg/mL. The TNT and F+TNT morphologies were verified using SEM and Raman spectra, respectively. The integrity of the cells in fibrin and the expression of the male and female germline stem cell markers Mvh/Ddx4, Dazl, and Plzf were assessed by immunofluorescence analysis, western blots, and real-time quantitative polymerase chain reaction (RT-qPCR) following multipotent mBMMSC culture in retinoic acid (RA) and F+TNT formation with RA and UV radiation, respectively. We demonstrate that a suitable two-dimensional (2D) scaffold can be obtained for germ-like cells derived from mBMMSCs through the use of F+TNT, UV radiation, and assisted reproductive technology (ART) in vitro maturation (IVM).