Protocol for the isolation, culture, and transfection of squid primary cells.

We present a protocol for the isolation, culture, and transfection of squid primary cells from various tissues and ages by including steps for squid dissection, cell isolation, maintenance in culture, and expression of exogenous genes. We also highlight life-stage tractability in Euprymna berryi squids for future aging studies. This protocol enables molecular- and cellular-level investigations into processes specific to squids, including their complex behaviors, rapid color change mechanisms, and RNA editing capabilities, with broader implications in basic cell physiology.