title: Real-time monitoring of specific oxygen uptake rates of embryonic stem cells in a microfluidic cell culture device.
abstract: 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.
authors.affiliation: Department of Biochemical Engineering, University College London, London, United Kingdom.
authors.forename: Alexandre
authors.initials: A
authors.lastname: Super
authors.affiliation: Department of Biochemical Engineering, University College London, London, United Kingdom.|Centre for Mathematics and Physics in the Life Sciences and Experimental Biology, University College London, London, United Kingdom.|Department of Computer Science, University College London, London, United Kingdom.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Jaccard
authors.affiliation: Department of Biochemical Engineering, University College London, London, United Kingdom.
authors.forename: Marco Paulo
authors.initials: MP
authors.lastname: Cardoso Marques
authors.affiliation: Department of Biochemical Engineering, University College London, London, United Kingdom.
authors.forename: Rhys Jarred
authors.initials: RJ
authors.lastname: Macown
authors.affiliation: Department of Computer Science, University College London, London, United Kingdom.
authors.forename: Lewis Donald
authors.initials: LD
authors.lastname: Griffin
authors.affiliation: Department of Biochemical Engineering, University College London, London, United Kingdom.
authors.forename: Farlan Singh
authors.initials: FS
authors.lastname: Veraitch
authors.affiliation: Department of Biochemical Engineering, University College London, London, United Kingdom. n.szita@ucl.ac.uk.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Szita
mesh_terms: D000818:Animals; D016466:CHO Cells; D002448:Cell Adhesion; D018929:Cell Culture Techniques / Q000295:instrumentation* / Q000379:methods; D049109:Cell Proliferation; D006224:Cricetinae; D003412:Cricetulus; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism; D007700:Kinetics; D051379:Mice; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation / Q000379:methods*; D008858:Microscopy, Phase-Contrast; D010100:Oxygen / Q000032:analysis* / Q000378:metabolism
chemical_list: D010100:Oxygen
keywords: Dissolved oxygen monitoring; Microfluidic cell culture device; Quantitative cell imaging; Specific oxygen uptake rate; Stem cells