Analysis of mitochondrial function and localisation during human embryonic stem cell differentiation in vitro.

Analysis of mitochondrial function and localisation during human embryonic stem cell differentiation in vitro.
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分析人类胚胎干细胞体外分化过程中线粒体的功能和定位。

DOI:
10.1371/journal.pone.0052214
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发表时间:
2012
期刊:
影响因子:
3.7
通讯作者:
Osborne GW
Osborne GW
中科院分区:
综合性期刊3区
文献类型:
--
作者:
Prowse AB;Chong F;Elliott DA;Elefanty AG;Stanley EG;Gray PP;Munro TP;Osborne GW

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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导致了一系列细胞类型的可靠生产和详细的分子特征。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改变的反应(如心肌细胞收缩),因此,线粒体表型可能在hESC分化的动态过程中发生变化。我们证明操纵线粒体生物发生改变中胚层承诺。为了研究hESCs早期谱系规范中的线粒体定位,我们开发了线粒体报告系KMEL2,其中编码绿色荧光蛋白(GFP)的序列靶向线粒体。分化成三种胚层的KMEL2系表明,这些分化后代的线粒体是GFP阳性的。因此,KMEL2 hESCs促进了线粒体在一系列细胞类型中的研究,重要的是,允许通过GFP标签对线粒体进行实时分析。
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.
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