Maintenance of Neurogenic Differentiation Potential in Passaged Bone Marrow-Derived Human Mesenchymal Stem Cells Under Simulated Microgravity Conditions.
Human mesenchymal stem cells (hMSCs) are considered to be able to adapt to environmental changes induced by gravity during cell expansion. In this study, we investigated neurogenic differentiation potential of passaged hMSCs under conventional gravity and simulated microgravity conditions. Immunostaining, quantitative real-time reverse transcription-polymerase chain reaction (qRT-PCR), and western blot analysis of neurogenic differentiation markers, neurofilament heavy (NF-H), and microtubule-associated protein 2 (MAP2) revealed that differentiated cells from the cells cultured under simulated microgravity conditions expressed higher neurogenic levels than those from conventional gravity conditions. The levels of NF-H and MAP2 in the cells from simulated microgravity conditions were consistent during passage culture, whereas cells from conventional gravity conditions exhibited a reduction of the neurogenic levels against an increase of their passage number. In growth culture, cells under simulated microgravity conditions showed less apical stress fibers over their nucleus with fewer cells having a polarization of lamin A/C than those under conventional gravity conditions. The ratio of lamin A/C to lamin B expression in the cells under simulated microgravity conditions was constant; however, cells cultured under conventional gravity conditions showed an increase in the lamin ratio during passages. Furthermore, analysis of activating H3K4me3 and repressive H3K27me3 modifications at promoters of neuronal lineage genes indicated that cells passaged under simulated microgravity conditions sustained the methylation during serial cultivation. Nevertheless, the enrichment of H3K27me3 significantly increased in the passaged cells cultured under conventional gravity conditions. These results demonstrated that simulated microgravity-coordinated cytoskeleton-lamin reorganization leads to suppression of histone modification associated with neurogenic differentiation capacity of passaged hMSCs.
['Koaykul|Chaiyong|C|', 'Kim|Mee-Hae|MH|', 'Kawahara|Yumi|Y|', 'Yuge|Louis|L|', 'Kino-Oka|Masahiro|M|']
D001854:Bone Marrow Cells D018929:Cell Culture Techniques D002454:Cell Differentiation D019070:Cell Lineage D049109:Cell Proliferation D003599:Cytoskeleton D018507:Gene Expression Regulation, Developmental D042421:Histone Code D006801:Humans D034904:Lamin Type A D059630:Mesenchymal Stem Cells D008869:Microtubule-Associated Proteins D016900:Neurofilament Proteins D055495:Neurogenesis D010012:Osteogenesis D011401:Promoter Regions, Genetic D018474:Weightlessness Simulation
neurogenic differentiation potential human mesenchymal stem cells simulated microgravity cytoskeleton histone methylation nuclear lamins