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选择最佳骨髓间充质干细胞通道进行肌萎缩侧索硬化症患者的干细胞治疗。

Selection of optimal passage of bone marrow-derived mesenchymal stem cells for stem cell therapy in patients with amyotrophic lateral sclerosis.

机构信息

Division of Molecular and Life Sciences, Hanyang University, Ansan, 426-791, Republic of Korea.

出版信息

Neurosci Lett. 2010 Mar 19;472(2):94-8. doi: 10.1016/j.neulet.2010.01.054. Epub 2010 Feb 1.

DOI:10.1016/j.neulet.2010.01.054
PMID:20117176
Abstract

Mesenchymal stem cells (MSCs) obtained from bone marrow (BM) are currently used as an alternative therapy in amyotrophic lateral sclerosis (ALS) patients. Selection of optimal passages of autologous BM-derived MSCs during long-term in vitro expansion is important for clinical trials in patients with ALS. We isolated and expanded MSCs from the BM of eight ALS patients to analyze the growth kinetics, differentiation potential, cellular surface antigen expression, karyotype modifications and secretion of various cytokines during long-term culture. The morphology and size of the cells changed from small and spindle-like cells to large and polygonal types in later passages. The growth rate of the MSCs was highest in the third passage, followed by a gradual decrease. There were no special modifications of cell surface antigens or the karyotype of the MSCs from the first to the tenth passage. MSCs in the fourth passage were differentiated into adipocytes, osteocytes and chondrocytes. When we analyzed the cultured media of MSCs at the third, fifth, seventh and ninth passages, IL-6, VEGF and IL-8 showed high expression, with more than 50pg/10,000 cells at these passages; however, their expression progressively decreased with additional passages. In addition, secretion of IL-15, GM-CSF, IL-10, PDGF-bb, G-CSF, IL-1beta, basic FGF and IFN-gamma gradually decreased over prolonged culture. We suggest that MSCs at earlier passages are more suitable for stem cell therapy in ALS patients because of their stability and more potent anti-inflammatory and neuroprotective properties.

摘要

间充质干细胞(MSCs)来源于骨髓(BM),目前被用作肌萎缩侧索硬化症(ALS)患者的替代治疗方法。在体外长期扩增过程中选择最佳的自体 BM 来源的 MSC 传代会对 ALS 患者的临床试验很重要。我们从 8 名 ALS 患者的 BM 中分离和扩增 MSC,以分析长期培养过程中的细胞生长动力学、分化潜能、细胞表面抗原表达、核型改变和各种细胞因子的分泌。细胞的形态和大小从小而纺锤形细胞变为大而多角形细胞,在后期传代中发生变化。MSC 的生长速度在第三代最高,随后逐渐下降。从第一代到第十代,MSC 的细胞表面抗原或核型没有特殊改变。第 4 代 MSC 分化为脂肪细胞、成骨细胞和软骨细胞。当我们分析第三代、第五代、第七代和第九代 MSC 培养物中的培养基时,IL-6、VEGF 和 IL-8 表现出高表达,在这些传代中超过 50pg/10,000 个细胞;然而,随着传代的增加,其表达逐渐下降。此外,随着培养时间的延长,IL-15、GM-CSF、IL-10、PDGF-bb、G-CSF、IL-1beta、碱性 FGF 和 IFN-γ的分泌逐渐减少。我们建议,由于早期传代的 MSC 具有稳定性和更强的抗炎和神经保护特性,因此更适合用于 ALS 患者的干细胞治疗。

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