Automated microfluidic cell culture of stem cell derived dopaminergic neurons

Abstract

Parkinson’s disease is a slowly progressive neurodegenerative disease characterised by dysfunction and death of selectively vulnerable midbrain dopaminergic neurons and the development of human in vitro cellular models of the disease is a major challenge in Parkinson’s disease research. We constructed an automated cell culture platform optimised for long-term maintenance and monitoring of different cells in three dimensional microfluidic cell culture devices. The system can be flexibly adapted to various experimental protocols and features time-lapse imaging microscopy for quality control and electrophysiology monitoring to assess cellular activity. Using this system, we continuously monitored the differentiation of Parkinson’s disease patient derived human neuroepithelial stem cells into midbrain specific dopaminergic neurons. Calcium imaging confirmed the electrophysiological activity of differentiated neurons and immunostaining confirmed the efficiency of the differentiation protocol. This system is the first example of an automated Organ-on-a-Chip culture and has the potential to enable a versatile array of in vitro experiments for patient-specific disease modelling.

Automation in the Life Science Research Laboratory

“…Life science laboratories often have a source of automation engineering expertise within their own institution in the form of engineering faculties. Both disciplines could benefit from increased interaction and discussion around laboratory automation, with examples of collaborating biomedicine and engineering departments producing innovative automated equipment (Kato et al, 2010;Kane et al, 2019). Collaboration at an educational level can be beneficial too.…”

Organ-on-a-chip: recent breakthroughs and future prospects

“…Peel et al [48] used automated systems to image multicellular OOACs, producing detailed cell phenotypes and statistical models for measurements. Kane et al [49] developed a cell system to monitor cells in a 3D microfluidic setting. These assays featured time-lapse imaging microscopy to assess cellular electrical activity through quality control.…”

Recent Developments in Microfluidic Technologies for Central Nervous System Targeted Studies

“…This model could be an economically efficient route to personalized drug discovery for PD [98]. Kane et al [99] established the first fully automated 3D brain-on-a-chip (termed "Pelican") for microfluidic cell culture and real-time monitorization through image acquisition. Using the Pelican, they differentiated PD patient stem cells into midbrain dopaminergic neurons.…”

“…The authors denote that this is a flexible cost-effective approach to not only model PD, but also for other diseases, enabling adaptation to various experimental protocols. Integration of new modules could expand even more the array of possible in vitro tests [99]. In a second study, Bolognin et al [100] also resorted to patient stem cells to generate a 3D microfluidic model with neurons carrying the LRRK2-G2019S mutation (responsible for autosomal-dominant PD).…”