A novel electro-chemotactic approach to impact the directional migration of transplantable retinal progenitor cells

Mishra, Shawn; Peña, Juan S.; Redenti, Stephen; Vázquez, Maribel

doi:10.1016/j.exer.2019.06.002

Cited by 14 publications

(16 citation statements)

References 56 publications

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“…Regenerative strategies rely strongly upon the local migration of replacement and/or supplemental cells (Mishra, Pena, Redenti, & Vazquez, ; Thakur et al, ), which requires detailed knowledge of cell migratory capacity and response(s). Although cell replacement therapies using terminal SCs remain elusive (Couve, Lovera, Suzuki, & Schmachtenberg, ), SC migration has been reported to aid axonal regeneration and synaptic SC–MN connections during transplantation of SCs, individually and in combination with different stem‐like cells and biomaterials (Ban et al, ; Hyung et al, ; Jones et al, ; Xia et al, ; Xie et al, ; Yang et al, ; Zeng et al, ; Zhang et al, ).…”

Section: Discussionmentioning

confidence: 99%

Neuronal substrates alter the migratory responses of nonmyelinating Schwann cells to controlled brain‐derived neurotrophic factor gradients

Singh

Robles

Vázquez

2020

J Tissue Eng Regen Med

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Neurodegeneration and dysfunction cause mobility impairment and/or paralysis in millions of adults, worldwide. Motor deficit and recovery in adults depend upon the plasticity of the neuromuscular junction (NMJ), a tripartite, biochemical synapse that transduces electrical impulses from the brain into voluntary contraction of skeletal muscle. Nonmyelinating Schwann cells (nmSCs) of the NMJ have been increasingly recognized as active synaptic partners with motor neurons and muscle and have become recent therapeutic targets for regeneration. nmSC synaptic transmission, plasticity, and growth are strongly modulated by brain-derived neurotrophic factor (BDNF), whose regenerative abilities have been explored through emerging biomaterials and tissue-engineered systems, as well as via clinical trials. Experimental models engineered to investigate integrated NMJ response(s) to local gradients of BDNF will both advance our understanding of key modulators of synaptic activity, postinjury, and aid in the development of NMJ-targeted, regenerative therapies to restore mobility. The current study examined the ability of nmSCs to respond to microfluidically controlled BDNF signaling upon different haptotactic substrates of motor neurons (MNs) and laminin adhesion coating. Tests seeding nmSCs sequentially with MNs illustrated that sequential seeding reported a fivefold increase in levels of tropomyosin receptor kinase B expression in response to BDNF signaling and a nearly fivefold increase in migration distance along BDNF gradients. By contrast, concurrent seeding of MNs and nmSCs upon laminin adhesion coating illustrated a difference in migration distance of less than one third-fold over control. Our findings are among the first to examine migratory responses of nmSCs for regenerative strategies and highlight the potential to restabilize NMJ synaptic activity by affecting nmSC behaviors through therapeutic BDNF and seeding with MNs.

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Section: Discussionmentioning

confidence: 99%

Neuronal substrates alter the migratory responses of nonmyelinating Schwann cells to controlled brain‐derived neurotrophic factor gradients

Singh

Robles

Vázquez

2020

J Tissue Eng Regen Med

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“…Although, these systems can create stable gradients, their applications have been limited since they often require extended times to establish a gradient due to the relatively low diffusivity in porous materials [30]. More complex designs that rely on sink and source channels connected with ladder-shaped microgroove arrays have been developed to overcome this problem [31][32][33][34]. In some of these designs, transient gradients can be established [35] in a short period of time; however, fabrication of these multi-depth [36] and multilayer channels [37] can be complex and require extensive usage of cleanroom facilities [38].…”

Section: Introductionmentioning

confidence: 99%

Chemotactic Responses of Jurkat Cells in Microfluidic Flow-Free Gradient Chambers

et al. 2020

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Gradients of soluble molecules coordinate cellular communication in a diverse range of multicellular systems. Chemokine-driven chemotaxis is a key orchestrator of cell movement during organ development, immune response and cancer progression. Chemotaxis assays capable of examining cell responses to different chemokines in the context of various extracellular matrices will be crucial to characterize directed cell motion in conditions which mimic whole tissue conditions. Here, a microfluidic device which can generate different chemokine patterns in flow-free gradient chambers while controlling surface extracellular matrix (ECM) to study chemotaxis either at the population level or at the single cell level with high resolution imaging is presented. The device is produced by combining additive manufacturing (AM) and soft lithography. Generation of concentration gradients in the device were simulated and experimentally validated. Then, stable gradients were applied to modulate chemotaxis and chemokinetic response of Jurkat cells as a model for T lymphocyte motility. Live imaging of the gradient chambers allowed to track and quantify Jurkat cell migration patterns. Using this system, it has been found that the strength of the chemotactic response of Jurkat cells to CXCL12 gradient was reduced by increasing surface fibronectin in a dose-dependent manner. The chemotaxis of the Jurkat cells was also found to be governed not only by the CXCL12 gradient but also by the average CXCL12 concentration. Distinct migratory behaviors in response to chemokine gradients in different contexts may be physiologically relevant for shaping the host immune response and may serve to optimize the targeting and accumulation of immune cells to the inflammation site. Our approach demonstrates the feasibility of using a flow-free gradient chamber for evaluating cross-regulation of cell motility by multiple factors in different biologic processes.

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“…A total of 40 cycles of 15 sec at 95 °C and 1 min at the annealing temperature of 60 °C were used to amplify the PCR product. The relative change in expression levels between the control and experimental conditions was calculated using the conventional ΔΔCT method, as used previously by our group [ 32 , 40 , 41 ]. Primer sequences are provided in Table 1 .…”

Section: Methodsmentioning

confidence: 99%

“…1020992-312, VWR, PA) and bonded to a chemically cleaned microscope slide (Nanostrip, VWR, MA) using ozone treatment. Reagents inserted into the source reservoir transport along the microchannel to the opposite reservoir via the well-established convective-diffusion model shown below:

where C (kg/m 3 )is the solution concentration, t (seconds) denotes time, ū (m/s) is bulk velocity, D (m 2 /s) is the diffusion coefficient, and x denotes position within the microchannel length [ 40 ].…”

Section: Methodsmentioning

confidence: 99%

VEGF Upregulates EGFR Expression to Stimulate Chemotactic Behaviors in the rMC-1 Model of Müller Glia

Peña

Vázquez

2020

Brain Sciences

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Progressive vision loss in adults has become increasingly prevalent worldwide due to retinopathies associated with aging, genetics, and epigenetic factors that damage the retinal microvasculature. Insufficient supply of oxygen and/or nutrients upregulates factors such as vascular endothelial growth factor (VEGF) and epidermal growth factor (EGF), which can induce abnormal angiogenesis and damage the structural arrangement of the retinal blood barrier (BRB). Müller glia (MG) regulate the diffusion of essential compounds across the BRB and respond to retinal insults via reactive gliosis, which includes cell hypertrophy, migration, and/or proliferation near areas of elevated VEGF concentration. Increasing concentrations of exogenous VEGF, upregulated by retinal pigmented epithelium cells, and endogenous epidermal growth factor receptor (EGF-R) stimulation in MG, implicated in MG proliferative and migratory behavior, often lead to progressive and permanent vision loss. Our project examined the chemotactic responses of the rMC-1 cell line, a mammalian MG model, toward VEGF and EGF signaling fields in transwell assays, and within respective concentration gradient fields produced in the glia line (gLL) microfluidic system previously described by our group. rMC-1 receptor expression in defined ligand fields was also evaluated using quantitative polymerase chain reaction (qPCR) and immunocytochemical staining. Results illustrate dramatic increases in rMC-1 chemotactic responses towards EGF gradient fields after pre-treatment with VEGF. In addition, qPCR illustrated significant upregulation of EGF-R upon VEGF pre-treatment, which was higher than that induced by its cognate ligand, EGF. These results suggest interplay of molecular pathways between VEGF and EGF-R that have remained understudied in MG but are significant to the development of effective anti-VEGF treatments needed for a variety of retinopathies.

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A novel electro-chemotactic approach to impact the directional migration of transplantable retinal progenitor cells

Cited by 14 publications

References 56 publications

Neuronal substrates alter the migratory responses of nonmyelinating Schwann cells to controlled brain‐derived neurotrophic factor gradients

Neuronal substrates alter the migratory responses of nonmyelinating Schwann cells to controlled brain‐derived neurotrophic factor gradients

Chemotactic Responses of Jurkat Cells in Microfluidic Flow-Free Gradient Chambers

VEGF Upregulates EGFR Expression to Stimulate Chemotactic Behaviors in the rMC-1 Model of Müller Glia

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