Nejvíce citovaný článek - PubMed ID 25194849
A sentinel protein assay for simultaneously quantifying cellular processes
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.
- Klíčová slova
- Cell line characterization, Mass spectrometry, Neural differentiation, Neural stem cell, Protein marker, Selected reaction monitoring,
- MeSH
- biologické markery MeSH
- buněčná diferenciace * MeSH
- buněčné linie MeSH
- buněčný rodokmen genetika MeSH
- hmotnostní spektrometrie MeSH
- imunohistochemie MeSH
- lidé MeSH
- nervové kmenové buňky cytologie metabolismus MeSH
- neuroglie MeSH
- neurony MeSH
- stanovení celkové genové exprese MeSH
- vývojová regulace genové exprese MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- biologické markery MeSH