Stability of human mesenchymal stem cells during in vitro culture: considerations for cell therapy
Stability of human mesenchymal stem cells during in vitro culture: considerations for cell therapy
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DOI：
10.1111/cpr.12002
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发表时间：
2013-02-01
影响因子：
8.5
通讯作者：
Abdelhay, E.
中科院分区：
文献类型：
--
作者：
Binato, R.;de Souza Fernandez, T.;Abdelhay, E.
人间充质干细胞的离体扩增和操作是免疫调节和再生细胞治疗的重要方法。尽管这些细胞显示出巨大的使用潜力，但与其整体性质相关的问题在该领域出现。需要在体外进行广泛的细胞数量扩增以获得足够的细胞数量以供使用，这会带来累积遗传和表观遗传异常的风险，从而可能导致散发性恶性细胞转化。在这项研究中，我们使用细胞遗传学分析、混合淋巴细胞反应、蛋白质组学和基因表达测定，在延长的培养时间内检查了源自骨髓的人类间充质干细胞，以确定培养物是否保留其在后续传代中使用的潜力。结果表明，这些细胞的体外培养物在第 4 代后表现出染色体变异，但其免疫调节功能和分化能力得以保持。在分子水平上，从第 5 代开始观察到变化，表明分化开始。总之，这些结果得出这样的假设：人类间充质干细胞培养物可成功用于细胞治疗，直至第 4 代。然而，使用更高代数的细胞必须根据具体情况进行分析。
Ex vivo expansion and manipulation of human mesenchymal stem cells are important approaches to immunoregulatory and regenerative cell therapies. Although these cells show great potential for use, issues relating to their overall nature emerge as problems in the field. The need for extensive cell quantity amplification in vitro to obtain sufficient cell numbers for use, poses a risk of accumulating genetic and epigenetic abnormalities that could lead to sporadic malignant cell transformation. In this study, we have examined human mesenchymal stem cells derived from bone marrow, over extended culture time, using cytogenetic analyses, mixed lymphocyte reactions, proteomics and gene expression assays to determine whether the cultures would retain their potential for use in subsequent passages. Results indicate that in vitro cultures of these cells demonstrated chromosome variability after passage 4, but their immunomodulatory functions and differentiation capacity were maintained. At the molecular level, changes were observed from passage 5 on, indicating initiation of differentiation. Together, these results lead to the hypothesis that human mesenchymal stem cells cultures can be used successfully in cell therapy up to passage 4. However, use of cells from higher passages would have to be analysed case by case.