Good Cell Culture Practices (GCCP) is of important relevance to in vitro toxicology. The European Society of Toxicology (ESTIV), the Center for Alternatives for Animal Testing (CAAT) and the In Vitro Toxicology Industrial Platform (IVTIP) joined forces to address by means of an ESTIV 2016 pre-congress session the different aspects and applications of GCCP. The covered aspects comprised new technological advances including stem cells, the development of an OECD guidance documents on Good In Vitro Method Practices, and the optimized implementation of Good Manufacturing Practices and Good Laboratory Practices for regulatory testing purposes. General discussions triggered by attendees raised the duality related to the difficulties in implementing such GCCP on an academic innovative research framework on one hand, and on the other hand, the need for such GCCP to ensure reproducibility and robustness of in vitro test methods for toxicity testing. Indeed, if good cell culture principles are critical to take into consideration for all uses of in vitro test methods for toxicity testing, the level of application of such principles may depend on the stage of development of the test method as well as on the applications of the test methods, i.e., academic innovative research vs. regulatory standardized test method.