Development of an N-Cadherin Biofunctionalized Hydrogel to Support the Formation of Synaptically Connected Neural Networks

O’Grady, Brian J.; Balotin, Kylie M.; Bosworth, Allison M.; McClatchey, P. Mason; Weinstein, Robert M.; Gupta, Mukesh Kumar; Poole, Kara; Bellan, Leon M.; Lippmann, Ethan S.

doi:10.1021/acsbiomaterials.0c00885

Cited by 24 publications

(17 citation statements)

References 88 publications

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“…iPSCs were passaged at 60-80% con uence using Versene (Gibco, Billings, MT) (38). iPSCs were differentiated into cortical glutamatergic neurons using previously described protocols (39) with minor modi cations (40). iPSCs were washed with 1x DPBS (Gibco) and incubated in Accutase (Gibco) for 3-5 minutes to obtain dissociated cells in suspension.…”

Section: Confocal Imaging and Quanti Cationsmentioning

confidence: 99%

Detection, Visualization and Quantification of Protein Complexes in Human Alzheimer’s Disease Brains using Proximity Ligation Assay

Romero-Fernández

Carvajal-Tapia

Prusky

et al. 2023

Preprint

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Examination of healthy and diseased human brain is essential to translational neuroscience. Protein-protein interactions play a pivotal role in physiological and pathological processes, but their detection is difficult, especially in aged and fixed human brain tissue. We used the proximity ligation assay (PLA) to broaden the range of molecular interactions assessable in-situ in human neuropathology. We adapted fluorescent in-situ PLA to detect ubiquitin-modified proteins in human brains with Alzheimer’s disease (AD), including approaches for the management of autofluorescence and quantification using a high-content image analysis system. We confirmed that hyperphosphorylated microtubule-associated protein tau (Serine202, Threonine205) aggregates were modified by ubiquitin and that phospho-tau-ubiquitin complexes were increased in hippocampal and frontal cortex regions in AD compared to non-AD brains. Overall, we refined PLA for use in human neuropathology, which has revealed a profound change in the distribution of ubiquitin in AD brain and its association with characteristic tau pathologies.

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Section: Confocal Imaging and Quanti Cationsmentioning

confidence: 99%

Detection, Visualization and Quantification of Protein Complexes in Human Alzheimer’s Disease Brains using Proximity Ligation Assay

Romero-Fernández

Carvajal-Tapia

Prusky

et al. 2023

Preprint

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“…Third, O'Grady et al 29 discussed biofunctionalized hydrogels in their research. Their findings suggest that the addition of N-cadherin extracellular peptide epitope to gelatin methacrylate (GelMA), resulting in a biomaterial termed GelMA-Cad supports the formation of synapses, hence, connected neural networks at the cellular and molecular levels.…”

Section: State Of the Art In Nanofabricating Neural Networkmentioning

confidence: 99%

Nanofabricating neural networks: Strategies, advances, and challenges

Lüttge

2022

Journal of Vacuum Science &Amp; Technology B

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Nanofabrication can help us to emulate natural intelligence. Forward-engineering brain gained enormous momentum but still falls short in human neurodegenerative disease modeling. Here, organ-on-chip (OoC) implementation of tissue culture concepts in microfluidic formats already progressed with the identification of our knowledge gap in toxicology and drug metabolism studies. We believe that the self-organization of stem cells and chip technology is a key to advance such complex in vitro tissue models, including models of the human nervous system as envisaged in this review. However, current cultured networks of neurons show limited resemblance with the biological functions in the real nervous system or brain tissues. To take full advantage of scaling in the engineering domain of electron-, ion-, and photon beam technology and nanofabrication methods, more research is needed to meet the requirements of this specific field of chip technology applications. So far, surface topographies, microfluidics, and sensor and actuator integration concepts have all contributed to the patterning and control of neural network formation processes in vitro. However, when probing the state of the art for this type of miniaturized three-dimensional tissue models in PubMed, it was realized that there is very little systematic cross-disciplinary research with biomaterials originally formed for tissue engineering purposes translated to on-chip solutions for in vitro modeling. Therefore, this review contributes to the formulation of a sound design concept based on the understanding of the existing knowledge and the technical challenges toward finding better treatments and potential cures for devastating neurodegenerative diseases, like Parkinson's disease. Subsequently, an integration strategy based on a modular approach is proposed for nervous system-on-chip (NoC) models that can yield efficient and informative optical and electronic NoC readouts in validating and optimizing these conceptual choices in the innovative process of a fast growing and exciting new OoC industry.

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“…Cortical glutamatergic neurons were generated from iPSCs as previously described 16 . Neurons were detached from plates by a 5-minute incubation with Accutase (Thermo Fisher Scientific), centrifuged, and pipetted into a Matrigel-coated 96-well plate at a density of 2x10 5 cells mL −1 .…”

Section: Direct Print Neuronal Toxicity Assaysmentioning

confidence: 99%