Isolation and culturing of primary mouse astrocytes for the analysis of focal adhesion dynamics

Baran E Güler; Jacek Krzysko; Uwe Wolfrum

doi:10.1016/j.xpro.2021.100954

Isolation and culturing of primary mouse astrocytes for the analysis of focal adhesion dynamics

STAR Protoc. 2021 Dec 8;2(4):100954. doi: 10.1016/j.xpro.2021.100954. eCollection 2021 Dec 17.

Authors

Baran E Güler¹, Jacek Krzysko¹, Uwe Wolfrum¹

Affiliation

¹ Institute of Molecular Physiology, Molecular Cell Biology, Johannes Gutenberg University of Mainz, Hanns-Dieter-Hüsch-Weg 17, 55128 Mainz, Germany.

Abstract

Primary astrocytes have gained attention as an important model for in vitro biological and biochemical research in the last decades. In this protocol, we describe a fast and cost-effective technique for isolating, culturing, and maintaining primary mouse astrocytes at ∼ 80% purity levels, which can be used in in vitro studies for migration and focal adhesion dynamics. In addition, we present an optimized transfection and manual quantification approach for focal adhesion analysis in fixed and living cells. For complete details on the use and execution of this protocol, please refer to Kusuluri et al. (2021).

Keywords: Cell Biology; Cell culture; Cell isolation; Microscopy; Neuroscience.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Astrocytes* / cytology
Astrocytes* / physiology
Cell Adhesion / physiology
Cell Culture Techniques / methods*
Cell Movement / physiology
Cell Separation / methods*
Cells, Cultured
Focal Adhesions / physiology*
Mice