Real-time monitoring of specific oxygen uptake rates of embryonic stem cells in a microfluidic cell culture device.
Real-time monitoring of specific oxygen uptake rates of embryonic stem cells in a microfluidic cell culture device.
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微流体细胞培养装置中胚胎干细胞的特定氧气吸收速率的实时监测。
DOI：
10.1002/biot.201500479
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发表时间：
2016-09
影响因子：
4.7
通讯作者：
Szita, Nicolas
中科院分区：
文献类型：
--
作者：
Super, Alexandre;Jaccard, Nicolas;Marques, Marco Paulo Cardoso;Macown, Rhys Jarred;Griffin, Lewis D.;Veraitch, Farlan Singh;Szita, Nicolas
关键词：
氧在干细胞生物学中作为信号分子和细胞能量代谢的指示剂起着关键作用。细胞氧动力学的定量，即比氧摄取率（苏尔）的测定，通常用于了解代谢变化。然而，目前测定贴壁细胞培养物中sOUR的方法依赖于细胞取样，这影响细胞表型。我们提出了真实的实时监测细胞生长的相差显微镜图像，呼吸使用光学传感器溶解氧。获得的中国仓鼠卵巢（CHO）和小鼠胚胎干细胞（mESC）培养物的原液和细胞氧浓度的时程数据成功证明了这种非侵入性和无标记方法。此外，我们通过将细胞暴露于线粒体抑制剂和解偶联剂，证实了对快速变化的培养条件的细胞反应的非侵入性检测。CHO和mESC的sOUR值分别为8 - 60阿莫尔cell-1 s-1和5 - 35阿莫尔cell-1 s-1。这些值与文献数据相比毫不逊色。非侵入性和真实的实时监测贴壁干细胞培养物的氧张力、细胞生长和sOUR的能力将对干细胞生物学和基于干细胞的疗法的未来研究具有重大益处。
Oxygen plays a key role in stem cell biology as a signaling molecule and as an indicator of cell energy metabolism. Quantification of cellular oxygen kinetics, i.e. the determination of specific oxygen uptake rates (sOURs), is routinely used to understand metabolic shifts. However current methods to determine sOUR in adherent cell cultures rely on cell sampling, which impacts on cellular phenotype. We present real‐time monitoring of cell growth from phase contrast microscopy images, and of respiration using optical sensors for dissolved oxygen. Time‐course data for bulk and peri‐cellular oxygen concentrations obtained for Chinese hamster ovary (CHO) and mouse embryonic stem cell (mESCs) cultures successfully demonstrated this non‐invasive and label‐free approach. Additionally, we confirmed non‐invasive detection of cellular responses to rapidly changing culture conditions by exposing the cells to mitochondrial inhibiting and uncoupling agents. For the CHO and mESCs, sOUR values between 8 and 60 amol cell−1 s−1, and 5 and 35 amol cell−1 s−1 were obtained, respectively. These values compare favorably with literature data. The capability to monitor oxygen tensions, cell growth, and sOUR, of adherent stem cell cultures, non‐invasively and in real time, will be of significant benefit for future studies in stem cell biology and stem cell‐based therapies.
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影响因子：
3.8
作者：
Bauwens, C;Yin, T;Zandstra, PW
通讯作者：
Zandstra, PW
影响因子：
5.2
作者：
Garita-Hernandez, Marcela;Diaz-Corrales, Francisco;Bhattacharya, Shomi S.
通讯作者：
Bhattacharya, Shomi S.
影响因子：
2.2
作者：
Gray, DR;Chen, S;Maiorella, BL
通讯作者：
Maiorella, BL
DOI：
10.1177/2211068214529288
发表时间：
2014-10
期刊：
Journal of laboratory automation
影响因子：
--
作者：
Jaccard N;Macown RJ;Super A;Griffin LD;Veraitch FS;Szita N
通讯作者：
Szita N
影响因子：
6.1
作者：
Fung, Wai-To;Beyzavi, Ali;Li, Hoi-Yeung
通讯作者：
Li, Hoi-Yeung