Work

Protocol for 3D Matrigel embedding of primary murine astrocytes for a physiologically relevant culture model

Year: 2025

Type: article

Abstract: Primary astrocyte 2D cultures often exhibit a phenotype that differs significantly from astrocytes in the brain. Here, we present a detailed protocol for culturing primary murine astrocytes within a 3D Matrigel environment, which better mimics the in vivo conditions. We describe a step-by-step guide for the isolation and preparation of primary striatal astrocytes, their embedding and cultivation in Matrigel, and the functional characterization of astrocytes grown in this 3D system. • Guided differentiation protocol of murine primary astrocytes • Steps for embedding primary murine astrocytes in Matrigel • Procedures for functional characterization of astrocytic uptake by live imaging Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. Primary astrocyte 2D cultures often exhibit a phenotype that differs significantly from astrocytes in the brain. Here, we present a detailed protocol for culturing primary murine astrocytes within a 3D Matrigel environment, which better mimics the in vivo conditions. We describe a step-by-step guide for the isolation and preparation of primary striatal astrocytes, their embedding and cultivation in Matrigel, and the functional characterization of astrocytes grown in this 3D system.

Source: STAR Protocols

DOI: 10.1016/j.xpro.2025.104165

Authors: Gurkirat Kaur, Maria Ester Salvucci, Veronica Giusti, Laura Civiero

Institutions: University of Padua, Ifremer, Nantes Université, IRCCS San Camillo Hospital

Language: English

Topic: Neuroscience and Neuropharmacology Research

Subfield: Cellular and Molecular Neuroscience

Field: Neuroscience

Domain: Life Sciences

SDG: Good health and well-being

Open Access status: gold

Estimated APC paid: $1,550