Real-time monitoring of specific oxygen uptake rates of embryonic stem cells in a microfluidic cell culture device

• Published in: Biotechnology journal 2016-09, Vol.11 (9), p.1179-1189
• Main Authors: Super, Alexandre, Jaccard, Nicolas, Cardoso Marques, Marco Paulo, Macown, Rhys Jarred, Griffin, Lewis Donald, Veraitch, Farlan Singh, Szita, Nicolas
• Format: Article
• Language: English
• Subjects: Animals; Cell Adhesion; Cell Culture Techniques - instrumentation; Cell Culture Techniques - methods; Cell Proliferation; CHO Cells; Cricetinae; Cricetulus; Dissolved oxygen monitoring; Embryonic Stem Cells - cytology; Embryonic Stem Cells - metabolism; Kinetics; Mice; Microfluidic Analytical Techniques - instrumentation; Microfluidic Analytical Techniques - methods; Microfluidic cell culture device; Microscopy, Phase-Contrast; Oxygen - analysis; Oxygen - metabolism; Quantitative cell imaging; Specific oxygen uptake rate; Stem cells
• ISSN: 1860-6768; 1860-7314
• DOI: 10.1002/biot.201500479

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. Oxygen is a critical molecule of the stem cell niche. Current approaches to measure specific oxygen uptake rate (sOUR) are disruptive to cell cultures. By combining optical oxygen sensors and state‐of‐the‐art quantitative imaging in a microfluidic cell culture device, the authors demonstrate for the first time real‐time quantification of sOUR in an uninterrupted adherent cell culture.