Survival and neurite outgrowth of rat cortical neurons in three-dimensional agarose and collagen gel matrices

O’Connor, Stephen M.; Stenger, David A.; Shaffer, Kara M.; Ma, Wu

doi:10.1016/s0304-3940(01)01769-4

Cited by 116 publications

(106 citation statements)

References 15 publications

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“…It has been shown that neurite extension on 3D gels depends upon the gel concentration and resultant mechanical stiffness (Balgude et al, 2001;O'Connor et al, 2001;Willits and Skornia, 2004), the presence of ECM molecules , and Ca 21 -dependent metalloproteinase from the growth cones (Pittman and Williams, 1989). Li and Folch (2005) showed that preferential growth of the axon tips is for a straight path, but also for a poly-D-lysine substrate over one with Matrigel.…”

Section: Discussionmentioning

confidence: 99%

Assessment of Axonal Growth into Collagen Nerve Guides Containing VEGF‐Transfected Stem Cells in Matrigel

Kempton

Gonzalez

Leven

et al. 2008

The Anatomical Record

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The early events associated with axonal growth into 10-mm nerve gaps were studied histologically in the rat sciatic nerve model to determine if the outgrowth of blood vessels, Schwann cells, and axons could be enhanced. In the first two experimental groups, collagen nerve guides were filled with either saline or Matrigel. Marrow-derived mesenchymal stem cells (MSCs) were added to Matrigel in two other groups, one of which contained cells transfected with VEGF (MSC/VEGF). After 21 days, the injury site was exposed, fixed, sectioned, and volume fractions of the conduit contents were determined by point counting. The bioresorbable collagen conduits appropriately guided the axons and vessels in a longitudinal direction. The volume fraction of axons was significantly greater in the group with saline when compared with all three groups with Matrigel. This measure had a significant positive correlation with actual counts of myelinated axons. The blood vessel volume fraction in the Matrigel group decreased compared with the saline group, but was restored in the MSC/VEGF group. All Matrigel groups had comparable cellularity and showed a distribution of residual Matrigel in acellular zones. The saline group, by contrast, sustained a network of delicate fibroblastic processes that compartmentalized the nerve and its natural matrix as it became infiltrated by axons as minifascicles. In conclusion, the reduction of axonal outgrowth in the Matrigel groups, when compared with the saline group, suggests that Matrigel may impede the early regenerative process even when enriched by the addition of MSCs or VEGF-transfected cells.

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Section: Discussionmentioning

confidence: 99%

Assessment of Axonal Growth into Collagen Nerve Guides Containing VEGF‐Transfected Stem Cells in Matrigel

Kempton

Gonzalez

Leven

et al. 2008

The Anatomical Record

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“…20,27,30,43 Models consisting of neurons distributed throughout a 3-D matrix material have previously been developed. 6,7,46,47,60 Dorsal root ganglia (DRG) extend neurites through hydrogel matrices in a manner dependent on the physical properties (e.g., agarose pore size 16 and stiffness 5 ), ligand concentration (e.g., collagen 59 and RGD peptides in fibrin 51 ), and substrate geometry. 62 Embryonic cortical neurons have been plated within 3-D matrices of collagen and various hydrogels (e.g., poly[N-(2-hydroxypropyl)-methacrylamide], 60 poly(acrylate), 46 and agarose 47 ).…”

Section: Introductionmentioning

confidence: 99%

“…6,7,46,47,60 Dorsal root ganglia (DRG) extend neurites through hydrogel matrices in a manner dependent on the physical properties (e.g., agarose pore size 16 and stiffness 5 ), ligand concentration (e.g., collagen 59 and RGD peptides in fibrin 51 ), and substrate geometry. 62 Embryonic cortical neurons have been plated within 3-D matrices of collagen and various hydrogels (e.g., poly[N-(2-hydroxypropyl)-methacrylamide], 60 poly(acrylate), 46 and agarose 47 ). Enhanced survival and neurite outgrowth was observed in collagen (0.4-0.5 mg/mL) as compared to hydrogels lacking ECM ligands, which produced varying degrees of cell viability and a paucity of neurite outgrowth, together indicating that growth and survival of primary cortical neurons are improved by specific cell-matrix interactions.…”

Section: Introductionmentioning

confidence: 99%

“…Enhanced survival and neurite outgrowth was observed in collagen (0.4-0.5 mg/mL) as compared to hydrogels lacking ECM ligands, which produced varying degrees of cell viability and a paucity of neurite outgrowth, together indicating that growth and survival of primary cortical neurons are improved by specific cell-matrix interactions. 46,47,60 Most in vitro models developed to study the response of neural cells to mechanical deformation utilize planar cell culture, and include mechanical stretch 19,23,54 and hydrodynamic shear stress 40,45 systems. Differences in cell morphology and the spatial distribution of cell-cell/cell-ECM interactions in planar vs. 3-D cultures may be important, especially in models of traumatic cellular injury as bulk deformation is translated to cells through physical coupling, which may be unrealistically represented in 2-D culture.…”

Section: Introductionmentioning

confidence: 99%

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Collagen-Dependent Neurite Outgrowth and Response to Dynamic Deformation in Three-Dimensional Neuronal Cultures

2007

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Abstract-In vitro models of brain injury that use thick 3-D cultures and control extracellular matrix constituents allow evaluation of cell-matrix interactions in a more physiologically relevant configuration than traditional 2-D cultures. We have developed a 3-D cell culture system consisting of primary rat cortical neurons distributed throughout thick (>500 lm) gels consisting of type IV collagen (Col) conjugated to agarose. Neuronal viability and neurite outgrowth were examined for a range of agarose (AG) percentages (1.0-3.0%) and initial collagen concentrations ([Col] i ; 0-600 lg/ mL). In unmodified AG, 1.5% gels supported viable cultures with significant neurite outgrowth, which was not found at lower (£1.0%) concentrations. Varying [Col] 30, 150, and 300 lg/mL, which supported cultures with similar baseline viability and neurite outgrowth. Conjugation of Col to AG also increased the complex modulus of the hydrogel. Following high rate deformation, neuronal viability significantly decreased with increasing [Col] i , implicating cell-matrix adhesions in acute mechanotransduction events associated with traumatic loading. These results suggest interrelated roles for matrix mechanical properties and receptor-mediated cell-matrix interactions in neuronal viability, neurite outgrowth, and transduction of high rate deformation. This model system may be further exploited for the elucidation of mechanotransduction mechanisms and cellular pathology following mechanical insult.

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“…Other groups have developed biosensors with cells cultured in a 3D-polymer matrix, including acrylamide derivatives, agarose, and collagen. [216][217][218] Neural progenitor cells entrapped within the collagen matrices forming 3D microspheres can give rise to neuronal progeny that are responsive to environmental toxicants. 218 This development can be further expanded to other cell types such as the immune cells and primary hepatocytes.…”

Section: Cell-based Biosensorsmentioning

confidence: 99%

Exploitation of physical and chemical constraints for three-dimensional microtissue construction in microfluidics

Choudhury

Iliescu

et al. 2011

Biomicrofluidics

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There are a plethora of approaches to construct microtissues as building blocks for the repair and regeneration of larger and complex tissues. Here we focus on various physical and chemical trapping methods for engineering three-dimensional microtissue constructs in microfluidic systems that recapitulate the in vivo tissue microstructures and functions. Advances in these in vitro tissue models have enabled various applications, including drug screening, disease or injury models, and cellbased biosensors. The future would see strides toward the mesoscale control of even finer tissue microstructures and the scaling of various designs for high throughput applications. These tools and knowledge will establish the foundation for precision engineering of complex tissues of the internal organs for biomedical applications.

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Survival and neurite outgrowth of rat cortical neurons in three-dimensional agarose and collagen gel matrices

Cited by 116 publications

References 15 publications

Assessment of Axonal Growth into Collagen Nerve Guides Containing VEGF‐Transfected Stem Cells in Matrigel

Assessment of Axonal Growth into Collagen Nerve Guides Containing VEGF‐Transfected Stem Cells in Matrigel

Collagen-Dependent Neurite Outgrowth and Response to Dynamic Deformation in Three-Dimensional Neuronal Cultures

Exploitation of physical and chemical constraints for three-dimensional microtissue construction in microfluidics

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