Wood Chelsea R, Juárez Esri H, Ferrini Francesco, Myint Peter, Innes John, Lossi Laura, Merighi Adalberto, Johnson William E B
Department of Biological Sciences, University of Chester, Parkgate Road, Chester, CH1 4BJ, UK.
Department of Veterinary Sciences, University of Turin, Largo Paolo Braccini 2, I-10095, Grugliasco, TO, Italy.
Biochem Biophys Rep. 2021 Mar 3;26:100976. doi: 10.1016/j.bbrep.2021.100976. eCollection 2021 Jul.
spinal cord slice cultures (SCSC) allow study of spinal cord circuitry, maintaining stimuli responses comparable to live animals. Previously, we have shown that mesenchymal stem/stromal cell (MSC) transplantation reduced inflammation and increased nerve regeneration but MSC survival was short-lived, highlighting that beneficial action may derive from the secretome. Previous studies of MSC conditioned medium (CM) have also shown increased neuronal growth. In this study, murine SCSC were cultured in canine MSC CM (harvested from the adipose tissue of excised inguinal fat) and cell phenotypes analysed via immunohistochemistry and confocal microscopy. SCSC in MSC CM displayed enhanced viability after propidium iodide staining. GFAP immunoreactivity was significantly increased in SCSC in MSC CM compared to controls, but with no change in proteoglycan (NG2) immunoreactivity. In contrast, culture in MSC CM significantly decreased the prevalence of βIII-tubulin immunoreactive neurites, whilst Ca transients per cell were significantly increased. These results contradict previous and reports of how MSC and their secretome may affect the microenvironment of the spinal cord after injury and highlight the importance of a careful comparison of the different experimental conditions used to assess the potential of cell therapies for the treatment of spinal cord injury.
脊髓切片培养（SCSC）能够对脊髓回路进行研究，保持与活体动物相当的刺激反应。此前，我们已经表明间充质干/基质细胞（MSC）移植可减轻炎症并促进神经再生，但MSC的存活时间较短，这突出表明有益作用可能源自其分泌组。先前对MSC条件培养基（CM）的研究也显示神经元生长增加。在本研究中，将小鼠SCSC培养于犬MSC CM（从切除的腹股沟脂肪的脂肪组织中收获）中，并通过免疫组织化学和共聚焦显微镜分析细胞表型。经碘化丙啶染色后，处于MSC CM中的SCSC显示出增强的活力。与对照组相比，处于MSC CM中的SCSC中GFAP免疫反应性显著增加，但蛋白聚糖（NG2）免疫反应性无变化。相反，在MSC CM中培养显著降低了βIII-微管蛋白免疫反应性神经突的发生率，而每个细胞的钙瞬变显著增加。这些结果与之前关于MSC及其分泌组如何影响脊髓损伤后微环境的报道相矛盾，并突出了仔细比较用于评估细胞疗法治疗脊髓损伤潜力的不同实验条件的重要性。
