模拟微重力下传代培养对间充质干细胞生物学特性影响的转录组学研究OACHSSCD

Objective To investigate the effects of prolonged passaging under simulated microgravity on critical biological properties of mesenchymal stem cells(MSCs),specifically their sternness,senescence,differentiation potential,proliferative capacity,and apoptotic regulation.Methods Mouse bone marrow-derived mesenchymal stem cells were continuously passaged in a simulated microgravity environment created by a 2D clinostat.Total RNA was extracted from cells at the 3rd(P3),6th(P6),and 9th(P9)passages in both simulated microgravity group and control group for transcriptomic sequencing and bioinformatic analysis.The expression of key functional markers in MSCs was further validated using qRT-PCR.Results Compared with the control group,the differentially expressed genes of P3,P6,and P9 cells cultured under simulated microgravity were significantly enriched in pathways related to immune response,signal transduction,senescence and apoptosis,and cell cycle.Specifically,the proliferative capacity of P3 and P6 cells was suppressed,while that of P9 cells was upregulated.Moreover,the expression levels of molecules associated with stemness maintenance,adipogenic differentiation,chondrogenic differentiation,and neurogenic differentiation were elevated across P3,P6,and P9 cells,accompanied by the increased expression of senescence-and apoptosis-related molecules(P＜0.05).Conclusion Long-term simulated microgravity can reshape the biological functions of MSCs,enhancing stemness,specific differentiation potential,and late-stage proliferation ability while activating senescence and apoptosis processes,thereby offering a novel perspective for optimizing the in vitro MSCs expansion system.