Evaluation of experimental parameters to model and assess long-term potentiation in vitro

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Examensarbete för masterexamen
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2019
Författare
Taha, Walid
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Impaired long-term potentiation (LTP), an essential neuronal process for learning and memory, is considered a pathological process during the course of several central nervous system (CNS) diseases. For instance, memory function is impaired in patients suffering from Alzheimer’s disease. The aim of this thesis is the identification of in vitro conditions to enable measuring LTP on a high throughput calcium-imaging platform. For this purpose, the high throughput platform Cellaxess Elektra, devised and produced by Cellectricon AB, was used to evaluate several in vitro parameters (cell culture media, cell density, electrical stimulation protocols) for their ability to be included in an assay measuring LTP in rodent neuronal cultures. Cell culture conditions were identified which showed spontaneous activity and evoked activity in the assay. However, in general, the average amplitude of spontaneous and evoked activity was observed to decrease over time. The cause for this will need to be further investigated as it compromised the analysis of LTP. The results also show that the cell culture medium “BrainPhysTM” is applicable to achieve healthy and electrophysiologically active primary cortical neurons in vitro. Since BrainPhys is a more physiologically relevant medium in comparison to previously applied cultivation media, a BrainPhys-based cultivation medium represents a promising medium for further investigation
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long-term potentiation, calcium imaging, high frequency stimulation
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