Restoring the quantity and quality of elderly human mesenchymal stem cells for autologous cell-based therapies.
Restoring the quantity and quality of elderly human mesenchymal stem cells for autologous cell-based therapies.
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DOI：
10.1186/s13287-017-0688-x
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发表时间：
2017-10-27
影响因子：
7.5
通讯作者：
Chen XD
中科院分区：
文献类型：
--
作者：
Block TJ;Marinkovic M;Tran ON;Gonzalez AO;Marshall A;Dean DD;Chen XD
退行性疾病是老龄化人口的主要公共卫生问题，间充质干细胞（MSC）在治疗许多此类疾病方面具有巨大潜力。然而，间充质干细胞的数量和质量随着年龄的增长而下降，限制了自体间充质干细胞治疗老年人群的潜在功效。根据国际细胞治疗学会 (ISCT) 的建议，使用标准细胞表面标记标准（CD73、CD90、CD105）从年轻和老年捐赠者获得人骨髓 (BM) 来源的 MSC 并进行表征。使用荧光激活细胞分选 (FACS) 根据大小和阶段特异性胚胎抗原 4 (SSEA-4) 表达将老年 MSC 群体分离为四个亚群，并使用评估 MSC 增殖、质量、形态、细胞内活性氧、β-半乳糖苷酶表达和三磷酸腺苷 (ATP) 含量的测定法将亚群与未分级的年轻和老年 MSC 进行比较。 ISCT 推荐的细胞表面标记物未能检测到年轻和老年 MSC 之间的任何差异。在这里，我们报告说，老年 MSC 体积较大，细胞内活性氧浓度和 β-半乳糖苷酶表达明显较高，而 ATP 和 SSEA-4 表达量较低。基于这些发现，通过 FACS 使用细胞大小和 SSEA-4 表达将老年 MSC 分为四个亚群。然后在组织培养塑料和 BM 衍生的细胞外基质 (BM-ECM) 上培养之前和之后对原始群体（年轻和老年 MSC）以及四个亚群进行表征。占原始老年MSC群体的8%左右的SSEA-4阳性小亚群表现出与年轻MSC相似的“年轻”表型。该老年亚群的生物活性受到未分级亲代群体产生的衰老相关因子的抑制。这些“年轻”细胞在“年轻微环境”（即来自年轻供体的BM细胞产生的BM-ECM）中被分离和扩增（传代3次）后，细胞数量增加了约17,000倍，达到3 × 109个细胞，并保留了其“年轻”表型。这些结果表明，从老年人群中获取大量高质量的自体间充质干细胞并建立个人干细胞库，连续输注“返老还童”的间充质干细胞来治疗与年龄相关的疾病是可行的。本文的在线版本 (doi:10.1186/s13287-017-0688-x) 包含补充材料，可供授权用户使用。
Degenerative diseases are a major public health concern for the aging population and mesenchymal stem cells (MSCs) have great potential for treating many of these diseases. However, the quantity and quality of MSCs declines with aging, limiting the potential efficacy of autologous MSCs for treating the elderly population. Human bone marrow (BM)-derived MSCs from young and elderly donors were obtained and characterized using standard cell surface marker criteria (CD73, CD90, CD105) as recommended by the International Society for Cellular Therapy (ISCT). The elderly MSC population was isolated into four subpopulations based on size and stage-specific embryonic antigen-4 (SSEA-4) expression using fluorescence-activated cell sorting (FACS), and subpopulations were compared to the unfractionated young and elderly MSCs using assays that evaluate MSC proliferation, quality, morphology, intracellular reactive oxygen species, β-galactosidase expression, and adenosine triphosphate (ATP) content. The ISCT-recommended cell surface markers failed to detect any differences between young and elderly MSCs. Here, we report that elderly MSCs were larger in size and displayed substantially higher concentrations of intracellular reactive oxygen species and β-galactosidase expression and lower amounts of ATP and SSEA-4 expression. Based on these findings, cell size and SSEA-4 expression were used to separate the elderly MSCs into four subpopulations by FACS. The original populations (young and elderly MSCs), as well as the four subpopulations, were then characterized before and after culture on tissue culture plastic and BM-derived extracellular matrix (BM-ECM). The small SSEA-4-positive subpopulation representing ~ 8% of the original elderly MSC population exhibited a “youthful” phenotype that was similar to that of young MSCs. The biological activity of this elderly subpopulation was inhibited by senescence-associated factors produced by the unfractionated parent population. After these “youthful” cells were isolated and expanded (three passages) on a “young microenvironment” (i.e., BM-ECM produced by BM cells from young donors), the number of cells increased ≈ 17,000-fold to 3 × 109 cells and retained their “youthful” phenotype. These results suggest that it is feasible to obtain large numbers of high-quality autologous MSCs from the elderly population and establish personal stem cell banks that will allow serial infusions of “rejuvenated” MSCs for treating age-related diseases. The online version of this article (doi:10.1186/s13287-017-0688-x) contains supplementary material, which is available to authorized users.
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影响因子：
7.5
作者：
Akiyama K;You YO;Yamaza T;Chen C;Tang L;Jin Y;Chen XD;Gronthos S;Shi S
通讯作者：
Shi S
影响因子：
13.6
作者：
Freund A;Orjalo AV;Desprez PY;Campisi J
通讯作者：
Campisi J
影响因子：
7.8
作者：
Nelson G;Wordsworth J;Wang C;Jurk D;Lawless C;Martin-Ruiz C;von Zglinicki T
通讯作者：
von Zglinicki T
影响因子：
64.8
作者：
Conboy, IM;Conboy, MJ;Rando, TA
通讯作者：
Rando, TA
影响因子：
18.2
作者：
Campisi J
通讯作者：
Campisi J