Efficient generation and cryopreservation of cardiomyocytes derived from human embryonic stem cells.
Efficient generation and cryopreservation of cardiomyocytes derived from human embryonic stem cells.
复制标题
DOI：
10.2217/rme.10.91
复制
发表时间：
2011-01
影响因子：
2.7
通讯作者：
Gold JD
中科院分区：
文献类型：
--
作者：
Xu C;Police S;Hassanipour M;Li Y;Chen Y;Priest C;O'Sullivan C;Laflamme MA;Zhu WZ;Van Biber B;Hegerova L;Yang J;Delavan-Boorsma K;Davies A;Lebkowski J;Gold JD
人胚胎干细胞（hESC）代表了治疗心力衰竭等疾病和用于药物筛选的新细胞来源。在这项研究中，我们的目标是促进心肌细胞从hESCs的有效生成相结合的控制未分化细胞的生长和心脏分化的特异性诱导目前的最佳技术。我们还旨在研究这些方法是否可扩展，以及分化的细胞是否可以冷冻保存。在没有条件培养基或饲养细胞的情况下维持hESC，并在无血清培养基中依次用激活素A和骨形态发生蛋白-4处理hESC。这导致分化成含有高百分比心肌细胞的细胞群。分化的细胞表达适当的心肌细胞标记物，并在培养中保持收缩性，大多数细胞显示工作室（心房和心室）型电生理特性。此外，细胞生长和分化过程适用于大型培养形式。此外，心肌细胞在冷冻保存后存活，并且在将冷冻保存的细胞移植到心肌梗死后的大鼠心脏中后检测到存活的心脏移植物。这些结果表明，高质量的心肌细胞可以有效地产生和冷冻保存使用hESC保持在无血清培养基中，这些细胞的应用，人类临床使用或药物发现的一个进步。
Human embryonic stem cells (hESCs) represent a novel cell source to treat diseases such as heart failure and for use in drug screening. In this study, we aim to promote efficient generation of cardiomyocytes from hESCs by combining the current optimal techniques of controlled growth of undifferentiated cells and specific induction for cardiac differentiation. We also aim to examine whether these methods are scalable and whether the differentiated cells can be cryopreserved. hESCs were maintained without conditioned medium or feeders and were sequentially treated with activin A and bone morphogenetic protein-4 in a serum-free medium. This led to differentiation into cell populations containing high percentages of cardiomyocytes. The differentiated cells expressed appropriate cardiomyocyte markers and maintained contractility in culture, and the majority of the cells displayed working chamber (atrial and ventricular) type electrophysiological properties. In addition, the cell growth and differentiation process was adaptable to large culture formats. Moreover, the cardiomyocytes survived following cryopreservation, and viable cardiac grafts were detected after transplantation of cryopreserved cells into rat hearts following myocardial infarctions. These results demonstrate that cardiomyocytes of high quality can be efficiently generated and cryopreserved using hESCs maintained in serum-free medium, a step forward towards the application of these cells to human clinical use or drug discovery.