Influence of in vitro biomimicked stem cell "niche' for regulation of proliferation and differentiation of human bone marrow-derived mesenchymal stem cells to myocardial phenotypes: serum starvation without aid of chemical agents and prevention of spontaneous stem cell transformation enhanced by the matrix environment
Influence of in vitro biomimicked stem cell "niche' for regulation of proliferation and differentiation of human bone marrow-derived mesenchymal stem cells to myocardial phenotypes: serum starvation without aid of chemical agents and prevention of spontaneous stem cell transformation enhanced by the matrix environment
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DOI：
10.1002/term.1754
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发表时间：
2016-01-01
影响因子：
3.3
通讯作者：
Suh, Hwal
中科院分区：
文献类型：
--
作者：
Kim, Jae Hyung;Shin, Sang-Hyun;Suh, Hwal
生态位对于防止细胞在体外扩增过程中发生的自发分化或衰老似乎很重要。在本研究中，我们发现人骨髓间充质干细胞（hBM-MSCs）在体外不经任何诱导处理的情况下，会经历与衰老相关的心肌谱系分化。这种现象发生在整个骨髓间充质干细胞群体中，与常规分化有很大不同，发生频率有限，并伴有形态变化，变成大而扁平的细胞，增殖受阻，这是骨髓间充质干细胞衰老的代表性迹象。通过对MSCs在多种培养条件下的培养，确定5-氮胞苷诱导处理与该现象无关，但在增殖严重受阻的血清饥饿条件下，导致衰老进程和心脏标志物上调。然而，在正常培养条件下，经过长时间的培养，MSCs逐渐形成心肌表型，形成异质群体。从临床应用的角度来看，为了公平和一致的结果，必须防止这种情况发生。因此，通过在常规可用的细胞外基质（ECM）上培养，为hBM-MSCs构建了仿生“生态位”。因此，ECM上的细胞恢复了纺锤形形态，增殖速度增加了两倍，并且在mRNA和蛋白质水平上显示心脏标志物的表达降低。综上所述，结果表明，在体外聚苯乙烯培养过程中，骨髓间充质干细胞衰老的进展可能通过心肌分化发生，这可以通过采用适当的ECM培养技术来克服。版权所有：John Wiley & Sons， Ltd。
Niche appears important for preventing the spontaneous differentiation or senescence that cells undergo during in vitro expansion. In the present study, it was revealed that human bone marrow-derived mesenchymal stem cells (hBM-MSCs) undergo senescence-related differentiation into the myocardial lineage in vitro without any induction treatment. This phenomenon occurred over the whole population of MCSs, much different from conventional differentiation with limited frequency of occurrence, and was accompanied by a change of morphology into large, flat cells with impeded proliferation, which are the representative indications of MSC senescence. By culturing MSCs under several culture conditions, it was determined that induction treatment with 5-azacytidine was not associated with the phenomenon, but the serum-starvation condition, under which proliferation is severely hampered, caused senescence progression and upregulation of cardiac markers. Nevertheless, MSCs gradually developed a myocardial phenotype under normal culture conditions over a prolonged culture period and heterogeneous populations were formed. In perspectives of clinical applications, this must be prevented for fair and consistent outcomes. Hence, the biomimetic 'niche' was constituted for hBM-MSCs by cultivating on a conventionally available extracellular matrix (ECM). Consequently, cells on ECM regained a spindle-shape morphology, increased in proliferation rate by two-fold and showed decreased expression of cardiac markers at both the mRNA and protein levels. In conclusion, the outcome indicates that progression of MSC senescence may occur via myocardial differentiation during in vitro polystyrene culture, and this can be overcome by employing appropriate ECM culture techniques. Copyright (c) 2013 John Wiley & Sons, Ltd.