年龄对猕猴骨髓间充质干细胞体外分化和细胞因子水平的影响

Age effects on in vitro differentiation and cytokine levels of macaque bone marrow mesenchymal stem cells

背景:间充质干细胞因其良好的生物学特性和广泛的应用前景而受到关注,但随着个体衰老和体外培养时间的增加,细胞逐渐呈现衰老的特征。非人灵长类动物与人具有相似的生物学特性,在动物模型和疾病治疗研究方面具有独特的优势。目的:分析不同年龄段猕猴骨髓间充质干细胞的增殖和分化情况,探讨年龄对骨髓间充质干细胞的影响及可能机制。方法:通过骨髓穿刺术采集3岁以内和20岁以上雄性猕猴的骨髓,分为幼年组和老年组,每组3只,分离骨髓间充质干细胞进行体外培养,观察其形态变化、增殖能力和分化能力,进行衰老相关的β-半乳糖苷酶染色,ELISA法和蛋白芯片检测细胞因子水平。结果与结论:随着年龄的增长,老年猕猴骨髓间充质干细胞的增殖、分化能力显著降低,衰老细胞显著增加,细胞因子白细胞介素1b、白细胞介素4、白细胞介素6、肿瘤坏死因子α和血管内皮生长因子水平显著降低,肝素结合碱性成纤维细胞生长因子和胎盘生长因子水平显著升高,说明骨髓间充质干细胞的增殖、分化、旁分泌等功能会因衰老而逐渐受到损害。

BACKGROUND:Mesenchymal stem cells attract extensive attention because of good biological characteristics and broad prospects,but the cells gradually show the characteristics of the aging with the increase of individual age or incubation time in vitro.Nonhuman primates have similar biological characteristics with human being,and have unique advantage in the animal model and disease treatment research.OBJECTIVE:To analyze the difference in proliferation and differentiation of bone marrow mesenchymal stem cells from macaques at different ages and to explore the effect of age on bone marrow mesenchymal stem cells and the possible mechanism.METHODS:Bone marrow samples from male macaques aged<3years and over20years were collected through bone marrow puncture,and divided into young group and elder group,with three macaques in each group.Then,bone marrow mesenchymal stem cells were isolated and cultured in vitro,and the morphological changes,proliferation and differentiation ability were observed.Age-related beta-galactosidase staining was performed,and protein microarray and ELISA were used to detect cytokine levels.RESULTS AND CONCLUSION:With age,the proliferation and differentiation of bone marrow mesenchymal stem cells from the elder macaques were reduced significantly,and the number of senescent cells increased significantly;the levels of interleukin-1b,interleukin-4,interleukin-6,tumor necrosis factorαand vascular endothelial growth factor were elevated obviously,the levels of heparin-binding basic fibroblast growth factor and placental growth factor were reduced.These findings indicate that the body's aging lead to the reduction in the proliferation,differentiation and cytokine secretion of bone marrow mesenchymal stem cells.