This review intends to provide a reflection regarding Directive 2010/63/EU, on animal welfare, and its application concerning cephalopod breeding and experimentation in aquaculture research. To do so, we gathered different perspectives of our group members, involving two cephalopod aquaculture researchers, an aquaculture fish production technician and a veterinary professional, the latter with no background on cephalopod research. The inclusion of this class in the animal welfare legislation; the definition of live cephalopods, stress, pain and suffering are revised according to the latest scientific knowledge. Considering the 15 year background that the Centre of Marine Sciences holds on the cultivation of the European cuttlefish, an evaluation of existing production protocols, aquaculture technology related research, ethics and cephalopod welfare are discussed. The application of anaesthesia, analgesia and euthanasia is discussed, bearing in mind the different procedures applied on a daily basis in aquaculture breeding and experimentation and the requirements of the new Directive. Finally, an overview of the above and progress on 3Rs (replacement of animals, reduction in number of animals and refinement of procedures) application to cephalopod welfare is presented.