Validation of the cardiosphere method to culture cardiac progenitor cells from myocardial tissue.
Validation of the cardiosphere method to culture cardiac progenitor cells from myocardial tissue.
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DOI：
10.1371/journal.pone.0007195
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发表时间：
2009-09-25
期刊：
影响因子：
3.7
通讯作者：
Marbán E
中科院分区：
文献类型：
--
作者：
Davis DR;Zhang Y;Smith RR;Cheng K;Terrovitis J;Malliaras K;Li TS;White A;Makkar R;Marbán E
至少有四个实验室已经证明，在原代培养中，内源性心脏前体细胞(CPC)可以直接从成人心脏组织中培养出来，作为心球或其后代(心球衍生细胞，CDCs)。事实上，疾病预防控制中心已经在心脏再生的临床试验中进行了测试。然而，基于不同方法的报告已经质疑了心圈战略产生CP的有效性。在这些报道中，心球被认为是心肌源性的，因为保留了心肌细胞，干细胞活性被认为是反映血液污染的指标。我们使用各种方法(包括遗传谱系追踪)来证明这两种人工产物都不适用于用现有方法培养的心球和CDCs，并进一步证明了CDCs的干细胞特征(即克隆性和多谱系潜力)。从人心内膜心肌活检组织(n = 160)、成年双转基因MercreMer-Z/EG小鼠(n = 6)、成年C57BL/6小鼠(n = 18)、成年绿色荧光蛋白+C57BL/6转基因小鼠(n = 3)、尤卡坦小型猪(n = 67)、成年米色小鼠(n = 8)和成年 = 大鼠(n = 80)中扩增CPC。胶原酶消化和工艺标准化提高了细胞产量；改变了培养基和特定动物品系的产量降低了。肝素化/器官逆行灌注不改变从心肌样本产生生长的能力。心肌样本的初始生长被浓缩为CPC(c-Kit+)、内皮细胞(CD31+、CD34+)和间充质细胞(CD90+)亚群。用MerCreMer-Z/EG转基因小鼠进行的谱系追踪表明，细胞突起中的心肌细胞不是生成CPC所必需的。大鼠树突状细胞是克隆性的，克隆的树突状细胞具有自发的多系分化潜能。这项研究表明，直接从心肌组织中培养和扩增CPC是简单、直接和可重复性的，当使用适当的技术时。
At least four laboratories have shown that endogenous cardiac progenitor cells (CPCs) can be grown directly from adult heart tissue in primary culture, as cardiospheres or their progeny (cardiosphere-derived cells, CDCs). Indeed, CDCs are already being tested in a clinical trial for cardiac regeneration. Nevertheless, the validity of the cardiosphere strategy to generate CPCs has been called into question by reports based on variant methods. In those reports, cardiospheres are argued to be cardiomyogenic only because of retained cardiomyocytes, and stem cell activity has been proposed to reflect hematological contamination. We use a variety of approaches (including genetic lineage tracing) to show that neither artifact is applicable to cardiospheres and CDCs grown using established methods, and we further document the stem cell characteristics (namely, clonogenicity and multilineage potential) of CDCs. CPCs were expanded from human endomyocardial biopsies (n = 160), adult bi-transgenic MerCreMer-Z/EG mice (n = 6), adult C57BL/6 mice (n = 18), adult GFP+ C57BL/6 transgenic mice (n = 3), Yucatan mini pigs (n = 67), adult SCID beige mice (n = 8), and adult Wistar-Kyoto rats (n = 80). Cellular yield was enhanced by collagenase digestion and process standardization; yield was reduced in altered media and in specific animal strains. Heparinization/retrograde organ perfusion did not alter the ability to generate outgrowth from myocardial sample. The initial outgrowth from myocardial samples was enriched for sub-populations of CPCs (c-Kit+), endothelial cells (CD31+, CD34+), and mesenchymal cells (CD90+). Lineage tracing using MerCreMer-Z/EG transgenic mice revealed that the presence of cardiomyocytes in the cellular outgrowth is not required for the generation of CPCs. Rat CDCs are shown to be clonogenic, and cloned CDCs exhibit spontaneous multineage potential. This study demonstrates that direct culture and expansion of CPCs from myocardial tissue is simple, straightforward, and reproducible when appropriate techniques are used.
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影响因子：
24
作者：
Li, Zongjin;Lee, Andrew;Huang, Mei;Chun, Hyung;Chung, Jaehoon;Chu, Pauline;Hoyt, Grant;Yang, Phillip;Rosenberg, Jarrett;Robbins, Robert C.;Wu, Joseph C.
通讯作者：
Wu, Joseph C.
影响因子：
20.1
作者：
Sohal, DS;Nghiem, M;Molkentin, JD
通讯作者：
Molkentin, JD
影响因子：
5.2
作者：
Andersen, Ditte Caroline;Andersen, Peter;Sheikh, Soren Paludan
通讯作者：
Sheikh, Soren Paludan
DOI：
10.1073/pnas.0706760104
发表时间：
2007-08-28
影响因子：
11.1
作者：
Bearzi, Claudia;Rota, Marcello;Anversa, Piero
通讯作者：
Anversa, Piero
影响因子：
158.5
作者：
Beltrami, AP;Urbanek, K;Anversa, P
通讯作者：
Anversa, P