Generation of cell-type-specific proteomes of neurodevelopment from human cerebral organoids

Abstract

Summary: Characterization of cerebral organoids has been challenging due to their heterogeneous nature. Here, we optimized a protocol to streamline the generation of FACS-purified cell populations from human cerebral organoids for proteomic analysis with liquid chromatography tandem mass spectrometry (LC-MS/MS). We describe the procedures for enzymatic dissociation of organoids into single-cell suspension, the generation of cell-type-specific lysates, peptide extraction, and proteomic analysis. This generalizable approach can be used to study temporal and cell-type-specific protein dynamics in developing cerebral organoids.For complete details on the use and execution of this protocol, please refer to Melliou et al. (2022). : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Keywords

• Cell isolation
• Flow cytometry/Mass cytometry
• Neuroscience
• Proteomics
• Organoids
• Mass spectrometry