Australian Institute for Bioengineering and Nanotechnology, The University of Queensland, St. Lucia, Australia.
PLoS One. 2012;7(12):e52214. doi: 10.1371/journal.pone.0052214. Epub 2012 Dec 19.
Human embryonic stem cell (hESC) derivatives show promise as viable cell therapy options for multiple disorders in different tissues. Recent advances in stem cell biology have lead to the reliable production and detailed molecular characterisation of a range of cell-types. However, the role of mitochondria during differentiation has yet to be fully elucidated. Mitochondria mediate a cells response to altered energy requirements (e.g. cardiomyocyte contraction) and, as such, the mitochondrial phenotype is likely to change during the dynamic process of hESC differentiation. We demonstrate that manipulating mitochondrial biogenesis alters mesendoderm commitment. To investigate mitochondrial localisation during early lineage specification of hESCs we developed a mitochondrial reporter line, KMEL2, in which sequences encoding the green fluorescent protein (GFP) are targeted to the mitochondria. Differentiation of KMEL2 lines into the three germ layers showed that the mitochondria in these differentiated progeny are GFP positive. Therefore, KMEL2 hESCs facilitate the study of mitochondria in a range of cell types and, importantly, permit real-time analysis of mitochondria via the GFP tag.
人类胚胎干细胞（hESC）衍生物有望成为多种不同组织疾病的可行细胞治疗选择。干细胞生物学的最新进展已经导致一系列细胞类型的可靠生产和详细的分子特征分析。然而，线粒体在分化过程中的作用尚未完全阐明。线粒体介导细胞对能量需求变化的反应（例如心肌细胞收缩），因此，线粒体表型在 hESC 分化的动态过程中可能会发生变化。我们证明了操纵线粒体生物发生会改变中胚层的承诺。为了研究 hESC 早期谱系特化过程中线粒体的定位，我们开发了一种线粒体报告系 KMEL2，其中编码绿色荧光蛋白（GFP）的序列靶向线粒体。KMEL2 系分化为三个胚层表明，这些分化后代中的线粒体是 GFP 阳性的。因此，KMEL2 hESC 有助于研究多种细胞类型中的线粒体，重要的是，通过 GFP 标签允许实时分析线粒体。
