Multi Electrode Array (MEA) technology allows to non-invasively record neural culture activity, overcoming several limits of traditional neuropharmacological and neurophysiological studies. To obtain reliable results and to ensure experiments reproducibility, it is necessary to maintain the cells in a controlled environment, which allows the proper growth of the culture. For this reason, MEAs recording setups need to be coupled with an environmental parameters control system, keeping CO2 and O2 concentration, temperature, humidity and pH within physiological ranges. In this work, we have validated a bench-top chamber prototype for long-term continuous and parallel recordings from multiple MEAs. We first validated the MEA acquisition chain through the comparison with a commercial gold-standard MEA recording system. Then, we tested a control unit based on environmental parameters sensors. To test the maintenance of optimal environmental conditions for cell cultures, long-term measurements of parameters were performed, alongside with measurements of culture medium evaporation rate and osmolarity. After demonstrating the capability of the setup to support cultures growth, neurophysiological trials were conducted on neuronal cell cultures inside the chamber. Through the comparison with the Benchmark system, we demonstrated that the custom benchtop system can monitor the cell electrical activity producing similar range of values for most of the features computed.
La tecnologia Multi Electrode Array (MEA) consente di registrare in modo non invasivo l'attività elettrofisiologica di colture neuronali, aggirando i diversi limiti degli approcci tradizionali. Per ottenere risultati affidabili e per garantire la riproducibilità degli esperimenti, è necessario mantenere le cellule in un ambiente controllato, che consenta la corretta crescita della cultura. Per questo motivo, i sistemi di acquisizione MEA devono essere accoppiati con un sistema di controllo dei parametri ambientali, mantenendo la concentrazione di CO2 e O2, temperatura, umidità e pH entro intervalli fisiologici. In questo lavoro, convalidiamo un prototipo di un sistema sperimentale per registrazioni a lungo termine continue e parallele su più MEA. Per prima cosa abbiamo convalidato la catena di acquisizione MEA attraverso il confronto con un sistema di registrazione commerciale. Poi, abbiamo testato un'unità di controllo basata su sensori di parametri ambientali. Per testare il mantenimento di un ambiente ottimale per colture cellulari, misurazioni a lungo termine sono state eseguite, oltre a misure di tasso di evaporazione di medium di coltura. Dopo aver dimostrato la capacità del setup di preservare la vitalità cellulare, studi neurofisiologici sono stati condotti su colture neuronali all'interno della camera di coltura. Abbiamo dimostrato che il setup personalizzato può monitorare l'attività elettrica della cellula restituendo valori di spiking e bursting estratti tramite algoritmi ad hoc confrontabili con quelli generati dalle registrazioni eseguite con il modello commerciale.
Development and validation of a culture chamber for neurophysiological trials on iPS neuron-like cells
PALAZZO, GIOVANNI
2017/2018
Abstract
Multi Electrode Array (MEA) technology allows to non-invasively record neural culture activity, overcoming several limits of traditional neuropharmacological and neurophysiological studies. To obtain reliable results and to ensure experiments reproducibility, it is necessary to maintain the cells in a controlled environment, which allows the proper growth of the culture. For this reason, MEAs recording setups need to be coupled with an environmental parameters control system, keeping CO2 and O2 concentration, temperature, humidity and pH within physiological ranges. In this work, we have validated a bench-top chamber prototype for long-term continuous and parallel recordings from multiple MEAs. We first validated the MEA acquisition chain through the comparison with a commercial gold-standard MEA recording system. Then, we tested a control unit based on environmental parameters sensors. To test the maintenance of optimal environmental conditions for cell cultures, long-term measurements of parameters were performed, alongside with measurements of culture medium evaporation rate and osmolarity. After demonstrating the capability of the setup to support cultures growth, neurophysiological trials were conducted on neuronal cell cultures inside the chamber. Through the comparison with the Benchmark system, we demonstrated that the custom benchtop system can monitor the cell electrical activity producing similar range of values for most of the features computed.File | Dimensione | Formato | |
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https://hdl.handle.net/10589/140238