biostudies-literatureDu WU.S. Department of Health & Human Services | NIH | National Cancer Institute (NCI)U.S. Department of Health & Human Services | NIH | National Institute of General Medical Sciences (NIGMS)NCI NIH HHSNIAMS NIH HHS30-42https://www.ebi.ac.uk/biostudies/studies/S-EPMC12917876biostudies-literatureUnknown23(1)Cell migration is a key cellular process that drives major developmental programs. To mimic and mechanistically understand cell migration in these different contexts, different assays have been developed. However, owing to the lack of practical guidelines, these different cell migration assays are often used interchangeably. This and the inherent dynamic nature of cell migration, which often requires sophisticated live-cell microscopy, may have caused cell migration to be notably less well understood than equally important cell functions, such as cell differentiation or proliferation. In this Review, we describe commonly used custom and commercial in vitro and in vivo cell migration assays and provide a comprehensive practical guide and decision tree outlining how to choose and implement aNature methodsSelecting the optimal cell migration assay: fundamentals and practical guidelines.PMC12917876UG3 CA275681UH3CA275681U54 AR081774U54 CA268083U54CA268083U54AR081774R35-GM157099UG3CA275681R01CA300052UH3 CA275681R01 CA300052Wu PHDu WPhillip JMForjaz AWirtz DNair PRfalseSelecting the optimal cell migration assay: fundamentals and practical guidelines.Cell migration is a key cellular process that drives major developmental programs. To mimic and mechanistically understand cell migration in these different contexts, different assays have been developed. However, owing to the lack of practical guidelines, these different cell migration assays are often used interchangeably. This and the inherent dynamic nature of cell migration, which often requires sophisticated live-cell microscopy, may have caused cell migration to be notably less well understood than equally important cell functions, such as cell differentiation or proliferation. In this Review, we describe commonly used custom and commercial in vitro and in vivo cell migration assays and provide a comprehensive practical guide and decision tree outlining how to choose and implement a2026-01-01T00:00:00Z2026 Jan2026-07-16T11:53:55.505Z2026-07-09T10:51:44.333ZS-EPMC129178764141334410.1038/s41592-025-02890-1