Agarose gel stiffness determines rate of DRG neurite extension in 3D cultures

Balgude, Amit; Yu, X; Szymanski, Agata; Bellamkonda, Ravi V.

doi:10.1016/s0142-9612(00)00350-1

Cited by 468 publications

(437 citation statements)

References 9 publications

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“…Interestingly, the rate of regeneration was 1-2 mm per week, but group differences at 2 or 6 weeks postoperative were absent. Neurites cultured in an agarose gel (1.25%) grow at a similar rate (0.25 mm/day) (Balgude et al, 2001). However, in most nerve graft models, the regeneration rate is 1-2 mm per day following an initial delay period of variable duration (Kerns et al, 1993).…”

Section: Discussionmentioning

confidence: 94%

“…Similarly, magnetically aligned laminin fibrils allow for better axonal growth when in a longitudinal compared with a horizontal orientation (Verdu et al, 2002). Gel density may also influence this directionality (Labrador et al, 1998;Balgude et al, 2001). The laminin concentration in Matrigel is high, ranging from 8 to 12 mg/mL according to the manufacturer's specifications.…”

Section: Discussionmentioning

confidence: 99%

“…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%

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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: 94%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

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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“…These studies 8,9 have shown that besides chemical cues, 3D matrix physical properties such as stiffness, 81 hydrophilicity, porosity, 29 pore size and void fraction 82,83 can affect cell morphology, attachment and function (Fig. 2).…”

Section: Importance Of Spatial Architecturementioning

confidence: 99%

Cell colonization in degradable 3D porous matrices

Lawrence

Madihally

2008

Cell Adhesion & Migration

170

144

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Cell colonization is an important in a wide variety of biological processes and applications including vascularization, wound healing, tissue engineering, stem cell differentiation and biosensors. During colonization porous 3D structures are used to support and guide the ingrowth of cells into the matrix. In this review, we summarize our understanding of various factors affecting cell colonization in three-dimensional environment. The structural, biological and degradation properties of the matrix all play key roles during colonization. Further, specific scaffold properties such as porosity, pore size, fiber thickness, topography and scaffold stiffness as well as important cell material interactions such as cell adhesion and mechanotransduction also influence colonization.

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“…Several recent studies have shown that soft substrates stimulated neurite extension and branching, but inhibited glial cell growth (Balgude et al, 2001;Engler et al, 2004;Flanagan et al, 2002;Georges et al, 2006;Lamoureux et al, 2002;Strassman et al, 1973). However, mechanisms of this regulation are not clear.…”

Section: Introductionmentioning

confidence: 99%

RPTPα is required for rigidity-dependent inhibition of extension and differentiation of hippocampal neurons

Kostić

Sap

Sheetz

2007

Journal of Cell Science

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Receptor-like protein tyrosine phosphatase α (RPTPα)-knockout mice have severe hippocampal abnormalities similar to knockouts of the Src family kinase Fyn. These enzymes are linked to the matrix-rigidity response in fibroblasts, but their function in neurons is unknown. The matrix-rigidity response of fibroblasts appears to differ from that of neuronal growth cones but it is unknown whether the rigidity detection mechanism or response pathway is altered. Here, we report that RPTPα is required for rigidity-dependent reinforcement of fibronectin (FN)-cytoskeleton bonds and the rigidity response in hippocampal neuron growth cones, like in fibroblasts. In control neurons, rigid FN surfaces inhibit neurite extension and neuron differentiation relative to soft surfaces. In RPTPα–/– neurons, no inhibition of extension and differentiation is found on both rigid and soft surfaces. The RPTPα-dependent rigidity response in neurons is FN-specific, and requires clustering of αvβ6 integrin at the leading edge of the growth cones. Further, RPTPα is necessary for the rigidity-dependent concentration of Fyn and p130Cas phosphorylation at the leading edge of the growth cone, like it is in fibroblasts. Although neurons respond to rigid FN surfaces in the opposite way to fibroblasts, we suggest that the mechanism of detecting FN rigidity is similar and involves rigidity-dependent RPTPα recruitment of Fyn.

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Agarose gel stiffness determines rate of DRG neurite extension in 3D cultures

Cited by 468 publications

References 9 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

Cell colonization in degradable 3D porous matrices

RPTPα is required for rigidity-dependent inhibition of extension and differentiation of hippocampal neurons

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