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bioRxiv · 10.1101/536664

Profiling embryonic stem cell differentiation by MALDI-MS: development of a reproducible and robust sample preparation workflow

Heap, R. E.·Segarra Fas, A.·Findlay, G.·Trost, M.
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Abstract

MALDI-TOF mass spectrometry (MS) is widely used to characterize and biotype bacterial samples, but a complimentary method for profiling of mammalian cells is still underdeveloped. Current approaches vary dramatically in their sample preparation methods and are not suitable for high-throughput studies. In this work, we present a universal workflow for mammalian cell MALDI-TOF MS analysis and apply it to distinguish ground-state naive and differentiating mouse embryonic stem cells (mESCs), which can be used as a model for drug discovery. We employed a systematic approach testing many parameters to evaluate how efficiently and reproducibly each method extracted unique mass features from four different human cell lines. This data enabled us to develop a unique mammalian cell MALDI-TOF workflow involving a freeze-thaw cycle, methanol fixing and CHCA matrix to generate spectra that yield maximum information and are highly reproducible. We applied our optimized workflow to distinguish naive and differentiating populations using multivariate analysis and reproducibly identifying unique features. Consequently, our MALDI-TOF MS profiling method enables identification of unique biomarkers and robust phenotyping of mESC differentiation. This method can in the future be applied to profile other cell types and expanded towards cellular MALDI-TOF MS screening assays.

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Heap, R. E., Segarra Fas, A., Findlay, G., Trost, M.. 2019-02-04. Profiling embryonic stem cell differentiation by MALDI-MS: development of a reproducible and robust sample preparation workflow. https://doi.org/10.1101/536664

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