A Novel Microfluidic Cell Co-culture Platform for the Study of the Molecular Mechanisms of Parkinson's Disease and Other Synucleinopathies

Abstract

Although, the precise molecular mechanisms underlying Parkinson's disease (PD) are still elusive, it is now known that spreading of alpha-synuclein (aSyn) pathology and neuroinflammation are important players in disease progression. Here, we developed a novel microfluidic cell-culture platform for studying the communication between two different cell populations, a process of critical importance not only in PD but also in many biological processes. The integration of micro-valves in the device enabled us to control fluid routing, cellular microenvironments, and to simulate paracrine signaling. As proof of concept, two sets of experiments were designed to show how this platform can be used to investigate specific molecular mechanisms associated with PD. In one experiment, naïve H4 neuroglioma cells were co-cultured with cells expressing aSyn tagged with GFP (aSyn-GFP), to study the release and spreading of the protein. In our experimental set up, we induced the release of the contents of aSyn-GFP producing cells to the medium and monitored the protein's diffusion. In another experiment, H4 cells were co-cultured with N9 microglial cells to assess the interplay between two cell lines in response to environmental stimuli. Here, we observed an increase in the levels of reactive oxygen species in H4 cells cultured in the presence of activated N9 cells, confirming the cross talk between different cell populations. In summary, the platform developed in this study affords novel opportunities for the study of the molecular mechanisms involved in PD and other neurodegenerative diseases.

Recent Developments in Microfluidic Technologies for Central Nervous System Targeted Studies

“…The second experiment entailed evaluating the impact of activated N9 microglia cells on the neuron-like H4 population. H4 cells co-cultured with the LPS-activated N9 cells presented almost 2-fold higher ROS levels than the controls, confirming the interplay and crosstalk between the two cell lines [101].…”

“…Fernandes et al [101] explored the molecular mechanisms involved in PD using a microfluidic chip. The device consisted of two culture chambers interconnected by three channels.…”

Patient-Derived Induced Pluripotent Stem Cell-Based Models in Parkinson’s Disease for Drug Identification

“…Such brain-on-a-chip platforms comprising miniaturized microfluidic perfusion systems that permit long-term growth in a format that is financially viable and has the potential of scaling up for launching high-throughput discovery campaigns, might pave the way for future fundamental discoveries and the development of more effective drugs [ 145 ]. Microfluidic devices have already been used to study the interaction of microglia and neurons in PD and to demonstrate neuronal internalization of αSyn fibrils before their propagation along the axons [ 146 , 147 , 148 ]. However, today, the most relevant formats are restricted to low-throughput applications awaiting their adaptation for automated screening on a large scale.…”

Lab-On-A-Chip for the Development of Pro-/Anti-Angiogenic Nanomedicines to Treat Brain Diseases

“…Some of the above devices were already evaluated for drug screening [32,34,36,38,40], and a few studied the effect of NPs [35,41] and NMs [39].Microfluidic technology has also been used to model brain vasculogenesis/angiogenesis [42][43][44][45][46], BBB [2,42,[47][48][49][50][51][52][53], brain tissues [54,55], and brain angiogenesis-related cellular events such as inflammation [47,50], cell migration [56], cell-cell interactions [57], etc., see Table 2. In addition, specific conditions, involving pathological-angiogenesis, such as brain tumors [46,[54][55][56]58], ischemic strokes [59], and neurodegenerative disorders including Alzheimer's disease (AD) [60], Parkinson's disease (PD) [61], and Huntington's disease (HD) [62], could also be created on-chip.In addition to this, LOCs have been developed for studying the biocompatibility, cellular uptake and transport of NMs [2,27], many of them focusing on brain angiogenesis [2,45,63]. Further, LOCs are also used to synthesis NMs [64].…”

“…Microfluidic technology has also been used to model brain vasculogenesis/angiogenesis [42][43][44][45][46], BBB [2,42,[47][48][49][50][51][52][53], brain tissues [54,55], and brain angiogenesis-related cellular events such as inflammation [47,50], cell migration [56], cell-cell interactions [57], etc., see Table 2. In addition, specific conditions, involving pathological-angiogenesis, such as brain tumors [46,[54][55][56]58], ischemic strokes [59], and neurodegenerative disorders including Alzheimer's disease (AD) [60], Parkinson's disease (PD) [61], and Huntington's disease (HD) [62], could also be created on-chip.…”