The annual congress of the SCMC (Societé Cerveau et Maldies Cérébraux Vasculaires – Brain and cerebrovascular diseases Society) will take place this Friday, may 28 2021 (virtual congress). On this occasion, our PhD student Véronique De Conto will present her PhD works, which concern the development of new cerebral in vitro model to better select drug candidate in drug discovery (abstract below).
Introduction: The pharmaceutical industry faces a very high failure rate in drug discovery, with only 10% of success. That points to a lack of relevance in preclinical models used upstream to select drug candidates, including in vitro models used for early screening. Among them, the SH-SY5Y cell line is commonly used for the assessment of neurotoxicity. Derived from a human neuroblastoma, SH-SY5Y cells can be differentiated into neurons using several methods. However, the consequence of the differentiation method on cell response is poorly documented.
Materials and methods: In this study, we have screened 24 differentiation methods on SH-SY5Y cells. After morphologic selection of the three most differentiating media (retinoic acid in 10% Fetal Bovine Serum (FBS), staurosporine in 1% FBS medium, and cAMP in B21-supplemented neurobasal medium), cells were analyzed for pan-neuronal and specific neuronal protein expression by fluorescent automated imaging. The response of SH-SY5Y to a set of compounds known as non-toxic, toxic but non-neurotoxic, or neurotoxic was examined in these culture conditions performed in 2D, and also in a 3D hyaluronic-based hydroscaffold which mimics the Extracellular Matrix (ECM).
Results: The expression of neuronal markers and the sensitivity to neurotoxic compounds varied according to the differentiation media used. The cAMP B21-supplemented neurobasal media led to the higher neuronal differentiation, and also the higher sensitivity to neurotoxic compounds. The culture in 3D in the hydroscaffold modified the cell response, with a lower sensitivity of cells cultured in 3D compared to the 2D culture.
Conclusion: The molecular and matricial microenvironment impacts on neurotoxic responses in vitro and thus should be considered carefully in research as well as in drug discovery.