Trettner Susanne, Seeliger Alexander, zur Nieden Nicole I
Department of Cell Therapy, Applied Stem Cell Technology Unit, Fraunhofer Institute for Cell Therapy and Immunology, Leipzig, Germany.
Methods Mol Biol. 2011;690:135-49. doi: 10.1007/978-1-60761-962-8_9.
While spontaneous differentiation is an undesired feature of expanding populations of embryonic stem cells, a variety of methods have been described for their intended differentiation into specialized cell types, such as the osteoblast or chondrocyte. Most commonly, differentiation initiation involves the aggregation of ESCs into a so-called embryoid body (EB), a sphere composed of approximately 15,000 differentiating cells. EB formation has been optimized through the years, for example through invention of the hanging drop protocol. Yet, it remains a highly laborious process. Here we describe the use of computer-controllable suspension bioreactors to form EBs in an automated and highly reproducible process and their subsequent differentiation along the osteoblast lineage. The development of the differentiating cells taken from bioreactor EBs to EBs formed in static control cultures through the hanging drop method will be compared.
虽然自发分化是胚胎干细胞扩增群体中不受欢迎的特征，但已经描述了多种方法来使其定向分化为特定细胞类型，如成骨细胞或软骨细胞。最常见的是，分化起始涉及胚胎干细胞聚集成所谓的胚状体（EB），这是一个由大约15,000个正在分化的细胞组成的球体。多年来，胚状体的形成已经得到优化，例如通过发明悬滴法。然而，这仍然是一个非常费力的过程。在这里，我们描述了使用计算机可控的悬浮生物反应器以自动化和高度可重复的过程形成胚状体，以及它们随后沿成骨细胞谱系的分化。将比较从生物反应器胚状体中获取的分化细胞到通过悬滴法在静态对照培养物中形成的胚状体的发育情况。
