Profiling embryonic stem cell differentiation by MALDI TOF mass spectrometry: development of a reproducible and robust sample preparation workflow

Heap, Rachel E.; Segarra-Fas, Anna; Blain, Alasdair; Findlay, Greg M.; Trost, Matthias

doi:10.1039/c9an00771g

Cited by 11 publications

(12 citation statements)

References 51 publications

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“…phosphate buffered saline, PBS) to remove residual culture medium and additives. PBS has been reported not to interfere with MALDI TOF MS significantly [49]. However, we observed that it may induce random quenching of ionization and decreased intensity of peaks.…”

Section: Intact Cell Maldi Tof Ms Of Hescsmentioning

confidence: 47%

Intact Cell Mass Spectrometry for Embryonic Stem Cell Biotyping

Vaňhara¹,

Moráň²,

Pečínka³

et al. 2021

Mass Spectrometry in Life Sciences and Clinical Laboratory

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Stem cells represent a unique cell type that is capable of self-renewal and differentiation into somatic cell types. Since the derivation of human embryonic stem cells and induced pluripotent stem cells, enormous potential has been recognized for disease modeling, drug development and regenerative medicine. Both embryonic stem cells and induced pluripotent stem cells possess the ability to differentiate into all three germ layers, hence they are naturally prone to respond to various differentiation stimuli. These inherent cellular fluctuations, which can result in risky phenotypic instability, must be addressed prior to introduction of these cells to human medicine, since they represent one of the major biosafety obstacles in the development of bio-industrial or clinical-grade stem cell cultures. Therefore, there is an ongoing need for novel robust, feasible and sensitive methods for determination and confirmation of the otherwise identical cells status, as well as for the detection of hidden divergences from their optimal state. A method of choice can be the intact cell mass spectrometry. Here we show how it can be applied in routine quality control of embryonic stem cell cultures.

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Section: Intact Cell Maldi Tof Ms Of Hescsmentioning

confidence: 47%

Intact Cell Mass Spectrometry for Embryonic Stem Cell Biotyping

Vaňhara¹,

Moráň²,

Pečínka³

et al. 2021

Mass Spectrometry in Life Sciences and Clinical Laboratory

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“…Several studies have noted the importance of uniform, homogeneous spots in MALDI–MS assays. [ 10,11,24 ] It is well known that DHB matrix can form inhomogeneous spots, with greatly varying intensities of analyte ions across the spot. This has prompted some authors to stop using it in assays, and others to employ matrix sprayers to prepare samples by spray‐coating rather than dried droplets.…”

Section: Resultsmentioning

confidence: 99%

“…This has prompted some authors to stop using it in assays, and others to employ matrix sprayers to prepare samples by spray‐coating rather than dried droplets. [ 10,11 ]…”

Section: Resultsmentioning

confidence: 99%

“…In recent years, MALDI–MS has emerged as a leading analytical tool in cell‐based drug discovery and profiling. [ 7–12 ] It is a versatile technology for the label‐free analysis of various classes of molecules, and is fast becoming established as a high‐throughput method for biochemical screening assays . [ 12–14 ] In non‐pharmacological applications, the utility of MALDI–MS for single cell metabolite analysis has been demonstrated, including in microcavity arrays.…”

Section: Introductionmentioning

confidence: 99%

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Fast Nanoliter‐Scale Cell Assays Using Droplet Microarray–Mass Spectrometry Imaging

et al. 2021

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In pharmaceutical research and development, cell‐based assays are primarily used with readout that rely on fluorescence‐based and other label‐dependent techniques for analysis of different cellular processes. Superhydrophobic–hydrophilic droplet microarrays (DMA) and matrix‐assisted laser desorption/ionization (MALDI) mass spectrometry (MS) have recently emerged as key technologies for miniaturized high‐throughput cell assays and for label‐free molecular high‐content drug profiling, respectively. Here, nanoliter‐scale cell assays are integrated on DMAs with MALDI–MS imaging (MALDI–MSI) approaches to a droplet microarray–mass spectrometry imaging (DMA–MSI) platform. Using A549 lung cancer cells, concentration‐response profiling of a pharmaceutical compound, the fatty acid synthase inhibitor GSK2194069, are demonstrated. Direct cell culture on DMAs enables combination of microscopy and high speed, high molecular content analysis using MALDI–MSI. Miniaturization of array spots down to 0.5 mm confining 40 nL droplets allows for MALDI imaging analysis of as few as ten cells per spot. Partial automation ensures a fast sample preparation workflow. Taken together, the integrated DMA–MSI platform that combines MALDI‐MSI, as a label‐free analytical readout, with the miniaturized droplet microarray platform is a valuable complement to high throughput cell‐based assays technologies.

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“…The use of MALDI-TOF-MS in the analysis of metabolites of low molecular weight compounds allows eliminating chromatographic separation [10]. MALDI-MS is widely used for the analysis of large compounds, such as proteins, peptides, and also whole bacteria cells [11][12][13]. Analysis of low molecular weight compounds by MALDI-MS is a current analytical challenge [14].…”

Section: Introductionmentioning

confidence: 99%

New Methodology for the Identification of Metabolites of Saccharides and Cyclitols by Off-Line EC-MALDI-TOF-MS

Sagandykova

Walczak-Skierska

Monedeiro

et al. 2020

IJMS

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A combination of electrochemistry (EC) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (off-line EC-MALDI-TOF-MS) was applied for determination of the studied biologically active compounds (D-glucose, D-fructose, D-galactose, D-pinitol, L-chiro-inositol, and myo-inositol) and their possible electrochemical metabolites. In this work, boron-doped diamond electrode (BDD) was used as a working electrode. MALDI-TOF-MS experiments were carried out (both in positive and negative ion modes and using two matrices) to identify the structures of electrochemical products. This was one of the first applications of the EC system for the generation of electrochemical products produced from saccharides and cyclitols. Moreover, exploratory data analysis approaches (correlation networks, hierarchical cluster analysis, weighted plots) were used in order to present differences/similarities between the obtained spectra, regarding the class of analyzed compounds, ionization modes, and used matrices. This work presents the investigation and comparison of fragmentation patterns of sugars, cyclitols, and their respective products generated through the electrochemistry (EC) process.

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Profiling embryonic stem cell differentiation by MALDI TOF mass spectrometry: development of a reproducible and robust sample preparation workflow

Abstract: A fast and robust sample preparation method for MALDI TOF MS analysis of whole mammalian cells and phenotypic profiling of mESCs.

Cited by 11 publications

References 51 publications

Intact Cell Mass Spectrometry for Embryonic Stem Cell Biotyping

Intact Cell Mass Spectrometry for Embryonic Stem Cell Biotyping

Fast Nanoliter‐Scale Cell Assays Using Droplet Microarray–Mass Spectrometry Imaging

New Methodology for the Identification of Metabolites of Saccharides and Cyclitols by Off-Line EC-MALDI-TOF-MS

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