A great deal is known about development in Sepia officinalis; however, much of this knowledge comes from animals incubated in laboratory conditions. Since cuttlefish are behaviorally plastic and known as embryos to perceive environmental stimuli from within the egg, we wondered if they are affected by incubation environment and thus whether laboratory-incubated cuttlefish exhibit natural behavior. We investigated the effects of incubation environment on hatchling size, defense and predation behavior in Sepia officinalis by comparing artificially-incubated hatchlings to naturally-incubated ones. Contrary to our expectations, no significant differences were apparent in hatchling size, disruptive body patterning and predation. A significant difference did exist between groups in one type of body patterning: Artificially-incubated hatchlings appeared to be better at producing a uniform body pattern than naturally-incubated individuals, possibly due to their prenatal experience with a homogeneous artificial substrate. This difference suggests some caution when interpreting experiments utilizing laboratory-incubated hatchlings, but overall, the limited effect of artificial incubation conditions demonstrated in this experiment bolsters confidence in previous behavioral results. These results are also promising for cuttlefish culture; eggs that would otherwise be lost as bycatch could instead be cultured artificially.