InVitreON: A compact and integrated platform for long-term neuronal culture incubation, recording and closed-loop stimulation

About This Grant

Neurological disorders constitute the largest global disease burden affecting ~43% of the world population, including but not limited to stroke, neonatal encephalopathy, dementia, Parkinson’s disease and epilepsy. Novel investigatory tools to study the fundamental basis for such nervous system malfunctions are critically needed to develop new therapies and prevention/rehabilitation strategies. Neurons are inherently different from most cell types; in that they are electrically active and form rich networks for communication and dynamic information storage. The proposed InVitreON culturing system will allow neuronal networks to be effectively investigated, modeled and used for screening potential therapies. It will significantly accelerate the mission to better Human health, while also allowing for fundamental studies to enhance scientific knowledge in behavioral areas pertaining to memory, pain, depression, anxiety and stress, among others. While in-vivo studies on Human and Animal subjects are used for clinically advanced stages in therapeutic development, a more ethical, well-controlled, and rapid approach has been to perform in-vitro studies of neuronal cultures. In-vitro culture systems allow live neuronal models of disease to be interrogated with a growing repertoire of molecular and cellular analysis techniques. Our InVitreON platform also allows for high-density neuro-electrophysiological functional studies. Over the past decades, in-vitro neuronal micro-electrode array (MEA) cultures have proven invaluable for disease modeling, neurotoxicity evaluations and drug screening. However, performing neuronal experiments with in-vitro MEAs has been plagued by several technical challenges that have discouraged use by typical neuroscientists (compared, for example, to optical microscopy). Current equipment solutions are complex, bulky, tedious to maintain, unreliable, inflexible, exhibit sub-optimal neuronal growth conditions and lack adequate contamination control. In addition, establishing closed-loop feedback via electrical, optical or chemical means, along with real-time raw data processing still requires deep engineering expertise, with no commercial product providing all these modalities. The focus of the proposed project is to disrupt this status quo, democratizing the use of neuronal in-vitro MEA platforms. This will be accomplished by Aim 1) Creating a single compact device that integrates live neuron culture incubation at physioxic (physiologically relevant) oxygen and carbon dioxide environments using mini-gas cartridges and capable of long-term MEA culture recording, with electromagnetic noise and contamination protection features. Aim 2) Demonstrating long- term >1 month live neuronal electrical recordings on primary rodent cortical, hippocampal neurons and Human iPSC derived neurons, while implementing stable automated media exchanges, and supported by integrated compute-rich single board processors for spike sorting and classification. Aim 3) Integrating flexible stimulation modalities including electrical, optical and chemical stimulation to expand the utility of the platform to support a diverse set of neuroscience experiments.