Targeted mass spectrometry for monitoring of neural differentiation

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Authors

SUCHA Rita KUBICKOVA Martina CERVENKA Jakub HRUSKA-PLOCHAN Marian BOHAČIAKOVÁ Dáša KEPKOVA VODICKOVA Katerina NOVAKOVA Tereza BUDKOVA Katerina SUSOR Andrej MARSALA Martin MOTLIK Jan KOVAROVA Hana VODICKA Petr

Year of publication 2021
Type Article in Periodical
Magazine / Source Biology Open
MU Faculty or unit

Faculty of Medicine

Citation
Web https://journals.biologists.com/bio/article/10/8/bio058727/271174/Targeted-mass-spectrometry-for-monitoring-of
Doi http://dx.doi.org/10.1242/bio.058727
Keywords Neural stem cell; Neural differentiation; Selected reaction monitoring; Mass spectrometry; Cell line characterization; Protein marker
Description Human multipotent neural stem cells could effectively be used for the treatment of a variety of neurological disorders. However, a defining signature of neural stem cell lines that would be expandable, non-tumorigenic, and differentiate into desirable neuronal/glial phenotype after in vivo grafting is not yet defined. Employing a mass spectrometry approach, based on selected reaction monitoring, we tested a panel of well-described culture conditions, and measured levels of protein markers routinely used to probe neural differentiation, i.e. POU5F1 (OCT4), SOX2, NES, DCX, TUBB3, MAP2, S100B, GFAP, GALC, and OLIG1. Our multiplexed assay enabled us to simultaneously identify the presence of pluripotent, multipotent, and lineage committed neural cells, thus representing a powerful tool to optimize novel and highly specific propagation and differentiation protocols. The multiplexing capacity of this method permits the addition of other newly identified cell type-specific markers to further increase the specificity and quantitative accuracy in detecting targeted cell populations. Such an expandable assay may gain the advantage over traditional antibody-based assays, and represents a method of choice for quality control of neural stem cell lines intended for clinical use.
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