Single Cerebral Organoid Mass Spectrometry of Cell-Specific Protein and Glycosphingolipid Traits

• Published in: Analytical chemistry (Washington) 2023-02, Vol.95 (6), p.3160-3167
• Main Authors: Nezvedová, Markéta, Jha, Durga, Váňová, Tereza, Gadara, Darshak, Klímová, Hana, Raška, Jan, Opálka, Lukáš, Bohačiaková, Dáša, Spáčil, Zdeněk
• Format: Article
• Language: English
• Subjects: Analytical chemistry; Astrocytes; Brain architecture; Cell Differentiation; Cell surface; Chemistry; Cultivation; Glial cells; Heterogeneity; Humans; Lipids; Mass Spectrometry; Mass spectroscopy; Nervous system; Neural Stem Cells; Neurogenesis; Neurological diseases; Neuronal-glial interactions; Neurons - metabolism; Organoids; Proteins; Radial glial cells; Reproducibility of Results; Stem cells; Workflow
• ISSN: 0003-2700; 1520-6882
• DOI: 10.1021/acs.analchem.2c00981

Cerebral organoids are a prolific research topic and an emerging model system for neurological diseases in human neurobiology. However, the batch-to-batch reproducibility of current cultivation protocols is challenging and thus requires a high-throughput methodology to comprehensively characterize cerebral organoid cytoarchitecture and neural development. We report a mass spectrometry-based protocol to quantify neural tissue cell markers, cell surface lipids, and housekeeping proteins in a single organoid. Profiled traits probe the development of neural stem cells, radial glial cells, neurons, and astrocytes. We assessed the cell population heterogeneity in individually profiled organoids in the early and late neurogenesis stages. Here, we present a unifying view of cell-type specificity of profiled protein and lipid traits in neural tissue. Our workflow characterizes the cytoarchitecture, differentiation stage, and batch cultivation variation on an individual cerebral organoid level.