Generation and enrichment of cerebellar GABAergic interneurons from human induced pluripotent stem cells and intracellular calcium measurements.
STAR Protoc
; 5(2): 102936, 2024 Jun 21.
Article
in En
| MEDLINE
| ID: mdl-38735042
ABSTRACT
GABAergic interneurons are inhibitory neurons of the CNS, playing a fundamental role in neural circuitry and activity. Here, we provide a robust protocol for the successful enrichment of human cerebellar GABAergic interneurons from human induced pluripotent stem cells (iPSCs) and measuring intracellular calcium transients. We describe in detail steps for culturing iPSCs; generating embryoid bodies; and differentiating and enriching for cerebellar GABAergic neurons (cGNs), with precise steps for their molecular characterization. We then detail the procedure for adeno-associated virus-mediated transduction of cGNs with genetically encoded calcium indicators, followed by intracellular calcium imaging and analyses. For complete details on the use and execution of this protocol, please refer to Pilotto et al.1.
Key words
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Cell Differentiation
/
Cerebellum
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Calcium
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Induced Pluripotent Stem Cells
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GABAergic Neurons
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Interneurons
Limits:
Humans
Language:
En
Journal:
STAR Protoc
Year:
2024
Document type:
Article
Affiliation country: