Neurons derived from P19 embryonic carcinoma cells as a platform for biosensor applications - optimisation and characterisation
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Abstract
P19 is a mouse-derived embryonal carcinoma cell line capable of differentiation
toward ectodermal, mesodermal and endodermal lineages and could thus be
differentiated into neurons. Different culture conditions
were tested to optimise and
increase the efficiency of neuronal differentiation since the population of P19-derived
neurons was reported to be heterogeneous with respect to the morphology and
neurotransmitters they synthesise. P19-derived neurons were cultured on
microelectrode arrays as cell aggregates and as dissociated cells. Improved neuronal
maturation was shown by the presence of microtubule associated protein 2,
neurofilament and synaptophysin formation when initiation of neuronal differentiation
was prolonged. High initial cell density cultures and coating of surfaces with
polyethylenimine-laminin further improved neuronal maturation of differentiated P19
cells. Increased spontaneous activities of the P19-derived neurons were
correspondingly recorded. Two to three hours recordings were performed between 17
and 25 days when extracellular signals were stabilised. It was found that P19-derived
neurons developed network properties as partially synchronised
network activities.
P19-derived neurons appeared to give inhomogenous response to the 2 major
neurotransmitters, -aminobutyric acid (GABA) and glutamate. The P19-derived
neuronal networks obtained from optimised protocol in this thesis were predominantly
GABAergic. The reproducible long term extracellular recordings performed showed
that neurons derived from P19 embryonal carcinoma cells could be applied as a model
for cell based biosensor in corporation with microelectrode arrays.