A Microfluidic Culture Platform to Assess Axon Degeneration

Yu, Yonghao; Hughes, Christopher C.W.; Deppmann, Christopher D.

doi:10.1007/978-1-0716-0585-1_7

Cited by 12 publications

(11 citation statements)

References 36 publications

(45 reference statements)

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“…The volume of each microwell was 1 mm 3 while the microwell diameter at the base was 900 μm. The disks were made from PDMS, a flexible material, permitting easy handling of the platform for hMO production, while being biologically compatible with the generation of neuronal tissue [61, 62].…”

Section: Resultsmentioning

confidence: 99%

Microfabricated disk technology: rapid scale up in midbrain organoid generation

Mohamed

Lépine

Lacalle‐Aurioles

et al. 2021

Preprint

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By providing a three-dimensional in vitro culture system with key features of the substantia nigra region in the brain, 3D neuronal organoids derived from human induced pluripotent stem cells (iPSCs) provide living neuronal tissue resembling the midbrain region of the brain. However, a major limitation of conventional brain organoid culture is that it is often labor-intensive, requiring highly specialized personnel for moderate throughput. Additionally, the methods published for long-term cultures require time-consuming maintenance to generate brain organoids in large numbers. With the increasing need for human midbrain organoids (hMOs) to better understand and model Parkinson′s disease (PD) in a dish, there is a need to implement new workflows and methods to both generate and maintain hMOs, while minimizing batch to batch variation. In this study, we developed a method with microfabricated disks to scale up the generation of hMOs. This opens up the possibility to generate larger numbers of hMOs, in a manner that minimizes the amount of labor required, while decreasing variability and maintaining the viability of these hMOs over time. Taken together, producing hMOs in this manner opens up the potential for these to be used to further PD studies.

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Section: Resultsmentioning

confidence: 99%

Microfabricated disk technology: rapid scale up in midbrain organoid generation

Mohamed

Lépine

Lacalle‐Aurioles

et al. 2021

Preprint

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“…Sympathetic neurons were dissected as described above and dissociated neurons were plated in microfluidic devices as previously described 36 , 37 . Cells were plated at a density of 14,000 cells per MFD.…”

Section: Methodsmentioning

confidence: 99%

Sympathetic neurons secrete retrogradely transported TrkA on extracellular vesicles

Mason

Keeler

Kabir

et al. 2023

Sci Rep

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Proper wiring of the peripheral nervous system relies on neurotrophic signaling via nerve growth factor (NGF). NGF secreted by target organs (i.e. eye) binds to the TrkA receptor expressed on the distal axons of postganglionic neurons. Upon binding, TrkA is internalized into a signaling endosome and retrogradely trafficked back to the soma and into the dendrites to promote cell survival and postsynaptic maturation, respectively. Much progress has been made in recent years to define the fate of the retrogradely trafficked TrkA signaling endosome, yet it has not been fully characterized. Here we investigate extracellular vesicles (EVs) as a novel route of neurotrophic signaling. Using the mouse superior cervical ganglion (SCG) as a model, we isolate EVs derived from sympathetic cultures and characterize them using immunoblot assays, nanoparticle tracking analysis, and cryo-electron microscopy. Furthermore, using a compartmentalized culture system, we find that TrkA derived from endosomes originating in the distal axon can be detected on EVs secreted from the somatodendritic domain. In addition, inhibition of classic TrkA downstream pathways, specifically in somatodendritic compartments, greatly decreases TrkA packaging into EVs. Our results suggest a novel trafficking route for TrkA: it can travel long distances to the cell body, be packaged into EVs, and be secreted. Secretion of TrkA via EVs appears to be regulated by its own downstream effector cascades, raising intriguing future questions about novel functionalities associated with TrkA+ EVs.

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“…These model systems fabricated from an inert polymer material such as poly(dimethylsiloxane) [ 55 , 56 ] comprise compartments and microgrooves of varying design, which spatially separate neuronal axons and soma [ 57 ]. Axons within these microfluidic devices extend into dedicated microgrooves, which allows the selective manipulation of axons and provides opportunities for the measurement of neurite growth response [ 58 ], somal transcription activity [ 56 ], and axonal degeneration [ 59 ]. In addition, these model systems have also been utilized to understand neuronal injury associated with amyotrophic lateral sclerosis [ 60 ], the overall dynamics of the neural network, and applications involving axonal biology, drug discovery, and regenerative medicine [ 61 ].…”

Section: Transection Injury Modelsmentioning

confidence: 99%

“…Another significant advancement in studying axonal injury is the adoption of microfluidic devices. Neurons cultured within microfluidic devices have been subjected to axonal injury by simply pushing fluid into intersecting microgrooves within the device [ 54 , 59 , 62 , 90 ]. Axonal stretching using a cantilever attached to a piezoelectric drive is another innovative approach to produce stretch-induced injury changes at the single axon level ( Figure 5 ).…”

Section: Axonal Stretch Injury Modelmentioning

confidence: 99%

A Brief Review of In Vitro Models for Injury and Regeneration in the Peripheral Nervous System

Varier

Raju

Madhusudanan

et al. 2022

IJMS

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Nerve axonal injury and associated cellular mechanisms leading to peripheral nerve damage are important topics of research necessary for reducing disability and enhancing quality of life. Model systems that mimic the biological changes that occur during human nerve injury are crucial for the identification of cellular responses, screening of novel therapeutic molecules, and design of neural regeneration strategies. In addition to in vivo and mathematical models, in vitro axonal injury models provide a simple, robust, and reductionist platform to partially understand nerve injury pathogenesis and regeneration. In recent years, there have been several advances related to in vitro techniques that focus on the utilization of custom-fabricated cell culture chambers, microfluidic chamber systems, and injury techniques such as laser ablation and axonal stretching. These developments seem to reflect a gradual and natural progression towards understanding molecular and signaling events at an individual axon and neuronal-soma level. In this review, we attempt to categorize and discuss various in vitro models of injury relevant to the peripheral nervous system and highlight their strengths, weaknesses, and opportunities. Such models will help to recreate the post-injury microenvironment and aid in the development of therapeutic strategies that can accelerate nerve repair.

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A Microfluidic Culture Platform to Assess Axon Degeneration

Cited by 12 publications

References 36 publications

Microfabricated disk technology: rapid scale up in midbrain organoid generation

Microfabricated disk technology: rapid scale up in midbrain organoid generation

Sympathetic neurons secrete retrogradely transported TrkA on extracellular vesicles

A Brief Review of In Vitro Models for Injury and Regeneration in the Peripheral Nervous System

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