Abstract
Tuberous sclerosis complex (TSC) is a neurodevelopmental disorder caused by deletions in the TSC1 or TSC2 genes that is associated with epilepsy in up to 90% of patients. Seizures are suggested to start in benign brain tumors, cortical tubers, or in the perituberal tissue making these tubers an interesting target for further research into mechanisms underlying epileptogenesis in TSC. Animal models of TSC insufficiently capture the neurodevelopmental biology of cortical tubers, and hence, human stem cell-based in vitro models of TSC are being increasingly explored in attempts to recapitulate tuber development and epileptogenesis in TSC. However, in vitro culture conditions for stem cell-derived neurons do not necessarily mimic physiological conditions. For example, very high glucose concentrations of up to 25 mM are common in culture media formulations. As TSC is potentially caused by a disruption of the mechanistic target of rapamycin (mTOR) pathway, a main integrator of metabolic information and intracellular signaling, we aimed to examine the impact of different glucose concentrations in the culture media on cellular phenotypes implicated in tuber characteristics. Here, we present preliminary data from a pilot study exploring cortical neuronal differentiation on human embryonic stem cells (hES) harboring a TSC2 knockout mutation (TSC2−/−) and an isogenic control line (TSC2+/+). We show that the commonly used high glucose media profoundly mask cellular phenotypes in TSC2−/− cultures during neuronal differentiation. These phenotypes only become apparent when differentiating TSC2+/+ and TSC2−/− cultures in more physiologically relevant conditions of 5 mM glucose suggesting that the careful consideration of culture conditions is vital to ensuring biological relevance and translatability of stem cell models for neurological disorders such as TSC.This article is part of the Special Issue “Proceedings of the 7th London-Innsbruck Colloquium on Status Epilepticus and Acute Seizures".
Highlights
Tuberous sclerosis complex (TSC) is a neurodevelopmental disorder, caused by mutations in the TSC1 or TSC2 genes, that is characterized by tumors in multiple organs [1]
We show preliminary data from a stem cell-based approach to the study of TSC highlighting the importance of using culture conditions that are as relevant as possible to the in vivo human physiology
Our data indicate that commonly used high glucose concentrations in vitro might mask important cellular phenotypes in a stem cell model of TSC
Summary
Tuberous sclerosis complex (TSC) is a neurodevelopmental disorder, caused by mutations in the TSC1 or TSC2 genes, that is characterized by tumors in multiple organs [1]. Stem cell-derived models are used to study a variety of different brain disorders including TSC [6,7,8], the pitfalls and key characteristics of these models are still to be fully discovered and described. Certain drawbacks, such as a considerable technical variability [9] and functional immaturity of derived neurons [10,11], are already well documented. Reliable neuronal differentiation is very dependent on cell culture media, which may support in vitro culture but not necessarily mimic human physiological conditions
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