We are pleased to participate to the 21st International Congress of the European Society of Toxicology In Vitro (ESTIV 2022), which will take place in Barcelona-Sitges, Spain, from 21st to 25th November 2022.
Our project Leader in in vitro pharmacology Véronique De Conto will present her poster entitled “In vitro microenvironment modifies the neurotoxic response of SH-SY5Y cells” (P-6a-8) during the session 6a: In vitro methods for safety testing of biopharmaceuticals/biotherapies/vaccines (Wednesday, November 23rd, 2022).
Introduction: Thirty-four percent of clinical trial safety failures are related to the brain, pointing to the lack of relevance of pre-clinical models for neurotoxicity assessment (Cook et al. 2014). Among these models, the SH-SY5Y neuroblastoma cell line is the most widely used for drug discovery (Kovalevich and Langford, 2013), but its differentiation into neurons in vitro is questioned. The objective of this study was to compared the SHSY5Y differentiation methods in order to develop a relevant in vitro model for neurotoxicity assessment, including both mature neurons organized in 3D and the Extracellular Matrix (ECM).
Materials & Methods: First, we screened the 24 major differentiation media described in the literature to differentiate these cells into neurons in 2D cell culture, and selected the 3 most differentiating conditions in terms of proliferation slowdown and neurite elongation: retinoic acid, staurosporine, and cyclic Adenosine Monophosphate (cAMP) combined to B21 supplement. Then, we compared their effects on SH-SY5Y cells cultivated in 3D in an hydroscaffold mimicking the ECM, by analyzing the cell proliferation, neuronal proteins expression, and sensitivity to compounds of known toxicity.
Results: Immunostainings showed that the three media decreased the cell proliferation, but only staurosporine and B21+cAMP media increased the neuronal marker expression in 3D. Both maturity and sensitivity of SH-SY5Y to the neurotoxic compound 6-hydroxydopamine were higher in B21+cAMP medium. However, cell sensitivity was lower than in 2D.
Conclusion: This work highlighted that the microenvironment of neurons, including the ECM and the soluble factors, can modify the neuronal response in vitro, and should thus be considered carefully in academic research and as early as possible in the drug discovery industrial process.
You can find the full published article here, or contact us for more information about our 3D cell culture technology BIOMIMESYS®, which mimics the ECM for more relevant in vitro models. Let’s replace animal testing by relevant human in vitro models!
Authors: Véronique De Conto1, Vaihere Cheung1, Gregory Maubon1, Zied Souguir1, Nathalie Maubon1, Elodie Vandenhaute1, Vincent Berezowski2 (1 HCS Pharma ; 2 Lille Neuroscience and Cognition, U1172)
References: Cook, D., Brown, D., Alexander, R., March, R., Morgan, P., Satterthwaite, G., and Pangalos, M.N. (2014). Lessons learned from the fate of AstraZeneca’s drug pipeline: a five-dimensional framework. Nat Rev Drug Discov 13, 419–431. https://doi.org/10.1038/nrd4309. Kovalevich, J., and Langford, D. (2013). Considerations for the use of SH-SY5Y neuroblastoma cells in neurobiology. Methods Mol. Biol. 1078, 9–21. https://doi.org/10.1007/978-1-62703-640-5_2.