Squid primary cell culture as a model system and experimental tool

Abstract

Squid primary cells from various tissues and ages are isolated, maintained in culture, and express exogenous genes. This protocol opens up numerous opportunities in molecular biology, neuroscience, and marine biology, enabling molecular and cellular-level investigations into processes specific to squids, including their complex behaviors, rapid color change mechanisms, RNA editing capabilities, with broader implications in basic cell physiology. We also describe procedures that harness life-stage tractability in Euprymna berryi squids towards future studies of aging in this model marine organism. Highlights- Squid primary cells are isolated from optic lobes, gills, eyes, and the skin - Cells from these tissues can be dissociated from any life stage of the squid - Isolated cells can express exogenous genes through mRNA transfections and are subject to live and/or fixed cell imaging - Specific details for optimized media conditions, trypsinization, plating, and passaging are included