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

ABSTRACT: 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).

SUBMITTER: Melliou S

PROVIDER: S-EPMC9597183 | biostudies-literature | 2022 Dec

REPOSITORIES: biostudies-literature

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

Melliou Sofia S Diamandis Phedias P

STAR protocols 20221021 4

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 gene ...[more]

PMID: 36313540

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