Abstract
A novel, non-invasive technique is reported for determining the numbers of cells in a culture by quantifying dimethyl sulphide (DMS) in the culture headspace as produced by the cellular enzymatic reduction of dissolved dimethyl sulphoxide (DMSO). Measured DMS concentrations, as performed using selected ion flow tube mass spectrometry (SIFT-MS), in the headspace of 2D and 3D cultures of four cell lines, viz. HEK293 (kidney), MG63 (bone), hepG2 (liver) and CALU-1 (lung), linearly correlate with starting cell number. Clear differences in the rates of production of DMS by the four cell types in both the 2D and 3D situations are seen. This novel analytical technique for cell enumeration offers a significant contribution to quality assessment across cell-based research and industry, including analysis of large scale culture systems, and for routine cell biology research.
Highlights
Mammalian cell cultures are involved in a range of applications, including the production of viral vaccines, recombinant proteins and cell therapies, as well as for various strands of biological research
This novel approach to cell number assessment obviously requires that the toxicity of dimethyl sulphoxide (DMSO) be addressed if the proposed method is to be utilized
A study by our group revealed no change in the viability of both primary human bone marrow-derived mesenchymal stem cells (MSCs) and hepG2 hepatocyte-like cells following 16 hours incubation in medium containing 0.1% DMSO.[21]
Summary
The unique cell assay described in this report is based on the enzymatic conversion of dimethyl sulphoxide (DMSO), one of the most commonly used amphipathic solvents,[11] to volatile dimethyl sulphide (DMS) in cell cultures by the actions of the ubiquitous enzyme methionine sulphoxide reductase A (MsrA) that has been identi ed in both the mitochondria and the cytosol of animal cells.[12]. This new method of cell enumeration is analogous to the aforementioned MTT assays except that the analysis can be performed non-invasively on a bulk population of cells
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