Abstract
Driven by an increasing number of studies demonstrating its relevance to a broad variety of disease states, the bioenergy production phenotype has been widely characterized at the bulk sample level. Its cell-to-cell variability, a key player associated with cancer cell survival and recurrence, however, remains poorly understood due to ensemble averaging of the current approaches. We present a technology platform for performing oxygen consumption and extracellular acidification measurements of several hundreds to 1,000 individual cells per assay, while offering simultaneous analysis of cellular communication effects on the energy production phenotype. The platform comprises two major components: a tandem optical sensor for combined oxygen and pH detection, and a microwell device for isolation and analysis of single and few cells in hermetically sealed sub-nanoliter chambers. Our approach revealed subpopulations of cells with aberrant energy production profiles and enables determination of cellular response variability to electron transfer chain inhibitors and ion uncouplers.
Highlights
Driven by an increasing number of studies demonstrating its relevance to a broad variety of disease states, the bioenergy production phenotype has been widely characterized at the bulk sample level
Increasing experimental evidence supports the notion of cell-to-cell variability as one of the key determinants in carcinogenesis and tumor progression in the context of clonal evolution mediated by complex interactions of cancer cells with their microenvironment[1,2,3,4]
We report on an integrated platform – the “Cellarium” – that enables combined characterization of oxygen consumption rate (OCR) and extracellular acidification rate (ECAR) of single cells with a throughput of up to 1,000 individual cells per assay
Summary
Driven by an increasing number of studies demonstrating its relevance to a broad variety of disease states, the bioenergy production phenotype has been widely characterized at the bulk sample level. We present a technology platform for performing oxygen consumption and extracellular acidification measurements of several hundreds to 1,000 individual cells per assay, while offering simultaneous analysis of cellular communication effects on the energy production phenotype. Our approach revealed subpopulations of cells with aberrant energy production profiles and enables determination of cellular response variability to electron transfer chain inhibitors and ion uncouplers. Underscoring the importance of bioenergy metabolism profiling are 2,231 published OCR/ECAR bulk cell studies performed since 2009 with the Seahorse platform alone. None of these technologies offer the sensitivity necessary to perform measurements at the single cell level. Our data revealed the existence of subpopulations of cells with both low OCR and ECAR under control conditions and in response to ETC inhibitors and proton uncouplers
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