Feeder layer- and serum-free culture of human embryonic stem cells
Feeder layer- and serum-free culture of human embryonic stem cells
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DOI：
10.1095/biolreprod.103.021147
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发表时间：
2004-03-01
影响因子：
3.6
通讯作者：
Itskovitz-Eldor, J
中科院分区：
文献类型：
--
作者：
Amit, M;Shariki, C;Itskovitz-Eldor, J
除了对人类早期发育研究的贡献外，人类胚胎干（hES）细胞也可用于基于细胞的疗法。传统上，这些细胞在小鼠胚胎成纤维细胞饲养层上培养，这使得它们能够在未分化状态下持续生长。然而，在人类治疗中使用 hES 细胞需要无动物的培养系统，其中避免暴露于小鼠逆转录病毒。在这项研究中，我们提出了一种新型的 hES 细胞无饲养层培养系统，该系统基于补充有 15% 血清替代品的培养基，以及包括转化生长因子 β1 (TGFbeta1)、白血病抑制因子、碱性成纤维细胞生长因子和纤连蛋白基质在内的生长因子组合。在这些条件下生长的人ES细胞在长时间培养后仍保持所有ES细胞特征，包括分化为三个胚胎胚层的代表性组织的发育潜力、无限且未分化的增殖能力以及正常核型的维持。这里介绍的培养系统有两个主要优点：1）应用明确的 hES 细胞培养系统；2）减少 hES 细胞与动物病原体的接触。本文报道的无饲养层培养系统旨在促进研究实践，并为 hES 细胞的未来临床应用提供更安全的替代方案。
In addition to their contribution to the research on early human development, human embryonic stem (hES) cells may also be used for cell-based therapies. Traditionally, these cells have been cultured on mouse embryonic fibroblast feeder layers, which allow their continuous growth in an undifferentiated state. However, the use of hES cells in human therapy requires an animal-free culture system, in which exposure to mouse retroviruses is avoided. In this study we present a novel feeder layer-free culture system for hES cells, based on medium supplemented with 15% serum replacement, a combination of growth factors including transforming growth factor beta1 (TGFbeta1), leukemia inhibitory factor, basic fibroblast growth factor, and fibronectin matrix. Human ES cells grown in these conditions maintain all ES cell features after prolonged culture, including the developmental potential to differentiate into representative tissues of the three embryonic germ layers, unlimited and undifferentiated proliferative ability, and maintenance of normal karyotype. The culture system presented here has two major advantages: 1) application of a well-defined culture system for hES cells and 2) reduced exposure of hES cells to animal pathogens. The feeder layer-free culture system reported here aims at facilitating research practices and providing a safer alternative for future clinical applications of hES cells.