Melatonin Treatment Improves Mesenchymal Stem Cells Therapy by Preserving Stemness during Long-term In Vitro Expansion.
Melatonin Treatment Improves Mesenchymal Stem Cells Therapy by Preserving Stemness during Long-term In Vitro Expansion.
复制标题
褪黑素治疗通过在长期体外扩增过程中保留干细胞来改善间充质干细胞治疗
作者：
Shuai Y;Liao L;Su X;Yu Y;Shao B;Jing H;Zhang X;Deng Z;Jin Y
间充质干细胞（MSCs）是组织再生和疾病治疗的有前途的候选者。然而，长期体外传代会导致MSCs的干性丧失，导致MSCs治疗失败。在这里，我们报告了一种基于褪黑素的策略来改善体外培养MSCs的细胞治疗。在四种具有抗衰老和干细胞保护特性的小分子（雷帕霉素、白藜芦醇、槲皮素和褪黑素）中，集落形成、增殖和成骨分化实验表明，褪黑素在长期传代后对大鼠骨髓间充质干细胞（BMMSCs）的自我更新和分化特性最有效。功能分析证实，褪黑素处理不影响培养1代或4代BMMSCs的集落形成、增殖和成骨分化，但在很大程度上阻止了体外培养15代BMMSCs自我更新和分化能力的下降。此外，异位成骨实验、临界尺寸颅骨缺损修复实验、骨质疏松治疗和实验性结肠炎治疗实验都有力地证明了褪黑素在体内保留了长期传代BMMSCs对骨再生和免疫治疗的治疗效果。从机制上讲，褪黑素通过激活抗氧化防御系统、抑制细胞衰老途径、维持干细胞调控基因的表达等发挥作用。综上所述，我们的研究结果表明，褪黑激素治疗有效地防止了BMMSCs在长期传代后的功能障碍和治疗失败，为提高BMMSCs在组织工程和细胞治疗中的应用提供了一种实用的策略。
Mesenchymal stem cells (MSCs) are promising candidates for tissue regeneration and disease treatment. However, long-term in vitro passaging leads to stemness loss of MSCs, resulting in failure of MSCs therapy. Here, we report a melatonin-based strategy to improve cell therapy of in vitro cultured MSCs. Among four small molecules with anti-aging and stem cell-protection properties (rapamycin, resveratrol, quercetin and melatonin), colony forming, proliferation, and osteogenic differentiation assay showed that melatonin was the most efficient to preserve self-renewal and differentiation properties of rat bone marrow MSCs (BMMSCs) after long-term passaging. Functional assays confirmed melatonin treatment did not affect the colony forming, proliferation and osteogenic differentiation of BMMSCs cultured for 1 or 4 passages, but largely prevented the decline of self-renew and differentiation capacity of BMMSCs cultured for 15 passages in vitro. Furthermore, heterotopic osteogenesis assay, critical size calvarial defects repair assay, osteoporosis treatment and experimental colitis therapy assay strongly certified that melatonin preserved the therapeutic effect of long-term passaged BMMSCs on bone regeneration and immunotherapy in vivo. Mechanistically, melatonin functioned by activating antioxidant defense system, inhibiting the pathway of cell senescence, and preserving the expression of gene governing the stemness. Taken together, our findings showed that melatonin treatment efficiently prevented the dysfunction and therapeutic failure of BMMSCs after long-term passaging, providing a practical strategy to improve the application of BMMSCs in tissue engineering and cytotherapy.