Donor age and long-term culture affect differentiation and proliferation of human bone marrow mesenchymal stem cells
Donor age and long-term culture affect differentiation and proliferation of human bone marrow mesenchymal stem cells
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DOI：
10.1007/s00277-012-1438-x
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发表时间：
2012-08-01
影响因子：
3.5
通讯作者：
Isik, Sevim
中科院分区：
文献类型：
--
作者：
Zaim, Merve;Karaman, Serap;Isik, Sevim
骨髓间充质干细胞（BM-hMSCs）是一种很有前途的基于细胞的治疗许多退行性疾病的方法。许多应用需要细胞扩增，并涉及治疗老年人群中发现的疾病和病症。因此，必须明确供体年龄和长期传代的影响。本研究探讨了供体年龄和长期传代对hMSCs形态、增殖能力、特性、中胚层分化能力和向神经源性谱系转分化能力的影响。来自儿童供体（0-12岁，n = 6）的细胞保持其成纤维细胞样形态直至更高的传代，并且比来自成人（25-50岁，n = 6）和老年（超过60岁，n = 6）供体的细胞增殖的数量更多。成脂、成骨和神经分化潜能随着年龄的增长而降低，而成软骨潜能没有变化。长期传代对各年龄段hMSCs的形态和增殖均有影响。随着传代次数的增加，细胞增殖率下降，细胞失去典型形态。神经标志物（β III微管蛋白和NSE）和拓扑异构酶II亚型在未分化的人骨髓间充质干细胞群体的表达水平进行了研究，通过逆转录聚合酶链反应分析。虽然神经标志物和拓扑II β表达水平增加，由于增加传代次数在成人hMSCs相比，儿童hMSCs，拓扑II α下降。这些结果表明，即使在高度标准化的培养条件下，供体年龄和长期传代对hMSC特性也有影响，这在基于干细胞的治疗之前应该考虑到。
Bone marrow-derived human mesenchymal stem cells (BM-hMSCs) represent a promising cell-based therapy for a number of degenerative conditions. Many applications require cell expansion and involve the treatment of diseases and conditions found in an aging population. Therefore, the effects of donor age and long-term passage must be clarified. In this study, the effects of donor age and long-term passage on the morphology, proliferation potential, characteristics, mesodermal differentiation ability, and transdifferentiation potential of hMSCs towards neurogenic lineage were evaluated. Cells from child donors (0-12 years, n = 6) maintained their fibroblast-like morphology up to higher passages and proliferated in a greater number than those from adult (25-50 years, n = 6) and old (over 60 years, n = 6) donors. Adipogenic, osteogenic, and neurogenic differentiation potential decreased with age, while chondrogenic potential did not change. Long-term passage affected the morphology and proliferation of hMSCs from all ages. With increasing passage number, proliferation rate decreased and cells lost their typical morphology. Expression levels of neural markers (beta III tubulin and NSE) and topo II isoforms in populations of nondifferentiated hMSCs were investigated by reverse transcription polymerase chain reaction analysis. While neural marker and topo II beta expression levels increased due to increasing passage number in adult hMSCs compared to child hMSCs, topo II alpha decreased in both. These results indicated that, even under highly standardized culture conditions, donor age and long-term passage have effects on hMSC characteristics, which should be taken into account prior to stem cell-based therapies.