Murine "Cardiospheres'' Are Not a Source of Stem Cells with Cardiomyogenic Potential
Murine "Cardiospheres'' Are Not a Source of Stem Cells with Cardiomyogenic Potential
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DOI：
10.1002/stem.72
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发表时间：
2009-01-01
期刊：
影响因子：
5.2
通讯作者：
Sheikh, Soren Paludan
中科院分区：
文献类型：
--
作者：
Andersen, Ditte Caroline;Andersen, Peter;Sheikh, Soren Paludan
最近的显著研究已经报道了具有心肌细胞潜能的克隆性推定心脏干细胞（CSCs）在离体培养期间从心脏组织活检中迁移，并且这些CSCs自组织成自发跳动的心脏球（CS）。这些数据提供了明确的承诺，即损伤的心脏组织可以通过使用自体CS衍生细胞的干细胞治疗来修复。通过进一步检查CS从原来的CS协议，使用免疫荧光，定量逆转录聚合酶链反应，和显微镜分析，我们在这里报告一个更平凡的结果：自发跳动的CS从新生大鼠可能包括污染心肌组织碎片。因此，过滤掉这些组织碎片导致CS没有心肌发生潜力。从新生小鼠获得的CS作为威尔斯孔获得与成年大鼠/小鼠相似的数据。此外，使用体外培养、荧光激活细胞分选和免疫荧光，我们证明这些CS是通过加塔-4（+）/胶原I+/α-平滑肌肌动蛋白（SMA）（+）/CD 45（-）细胞的细胞聚集产生的，而不是通过克隆细胞生长产生的。与此相反，我们发现先前提出的CS形成细胞，被称为相位亮细胞，是加塔-4（-）/胶原I-/α-SMA（-）/CD 45（+），并且不能自己形成CS。从表型上看，CS细胞很大程度上类似于成纤维细胞，并且它们缺乏心肌细胞和内皮细胞分化潜力。我们的数据表明，小鼠CS模型是不适合作为具有心肌细胞潜能的CSC的来源，这一结果与先前发表的数据相反。因此，我们建议，在启动人体试验之前，应进一步表征人类CS的表型和分化潜力。干细胞2009; 27：1571-1581
Recent remarkable studies have reported that clonogenic putative cardiac stem cells (CSCs) with cardiomyogenic potential migrate from heart tissue biopsies during ex vivo culture, and that these CSCs self-organize into spontaneously beating cardiospheres (CSs). Such data have provided clear promise that injured heart tissue may be repaired by stem cell therapy using autologous CS-derived cells. By further examining CSs from the original CS protocol using immunofluorescence, quantitative reverse transcription-polymerase chain reaction, and microscopic analysis, we here report a more mundane result: that spontaneously beating CSs from neonatal rats likely consist of contaminating myocardial tissue fragments. Thus, filtering away these tissue fragments resulted in CSs without cardiomyogenic potential. Similar data were obtained with CSs derived from neonatal mice as wells as adult rats/mice. Additionally, using in vitro culture, fluorescence-activated cell sorting, and immunofluorescence, we demonstrate that these CSs are generated by cellular aggregation of GATA-4(+)/collagen I+/alpha-smooth muscle actin (SMA)(+)/CD45(-) cells rather than by clonal cell growth. In contrast, we found that the previously proposed CS-forming cells, dubbed phase bright cells, were GATA-4(-)/collagen I-/alpha-SMA(-)/CD45(+) and unable to form CSs by themselves. Phenotypically, the CS cells largely resembled fibroblasts, and they lacked cardiomyogenic as well as endothelial differentiation potential. Our data imply that the murine CS model is unsuitable as a source of CSCs with cardiomyogenic potential, a result that is in contrast to previously published data. We therefore suggest, that human CSs should be further characterized with respect to phenotype and differentiation potential before initiating human trials. STEM CELLS 2009; 27: 1571-1581