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Isolation and Maintenance of Murine Embryonic Striatal Neurons.

Bio-protocol (2018-04-20)
Luana Naia, A Cristina Rego
ABSTRACT

Primary cultures of murine striatal neurons are widely used to explore cellular mechanisms in neurobiology, including brain diseases. Here we describe a detailed and standardized protocol to dissect and culture embryonic murine striatal neurons GABA-positive/DARPP-32-positive for 12 days in vitro, when they show good neuronal cell connectivity and the presence of dendritic spines, which reflects the maturation of the network.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Anti-GABA antibody produced in rabbit, affinity isolated antibody, buffered aqueous solution
Sigma-Aldrich
HEPES, ≥99.5% (titration)
Sigma-Aldrich
Trypsin inhibitor from Glycine max (soybean), lyophilized powder
Sigma-Aldrich
5-Fluoro-2′-deoxyuridine, thymidylate synthase inhibitor
Sigma-Aldrich
Deoxyribonuclease I from bovine pancreas, Type IV, lyophilized powder, ≥2,000 Kunitz units/mg protein
Sigma-Aldrich
Trypsin from porcine pancreas, Type IX-S, lyophilized powder, 13,000-20,000 BAEE units/mg protein