Laminin oligopeptide derivatized agarose gels allow three‐dimensional neurite extension in vitro

Abstract

The phenotypic expression of various neural cells is influenced by extracellular matrix (ECM) molecules. This study aims to develop a three-dimensional gel tailored to support neurite extension from neural cells. Laminin-derived (LN) oligopeptides CDP-GYIGSR, a 19-mer IKVAV containing sequence, GRGDSP, a cocktail of the three aforementioned LN peptides (PEPMIX), and a control peptide sequence GGGGG were covalently linked to an agarose hydrogel backbone using the bi-functional coupling agent 1'1, carbonyldiimidazole. Embryonic day 9 chick DRGs and PC12 cells were suspended in three dimensions in underivatized and derivatized agarose gels and neurite extension was analyzed. Agarose gels derivatized with CDPGYIGSR and PEPMIX enhanced neurite outgrowth from DRGs while GRGDSP and IKVAV derivatized gels inhibited neurite extension when compared to underivatized agarose gels. The IKVAV derivatized gels significantly enhanced neurite outgrowth from PC12 cells in comparison to underivatized and other LN peptide derivatized gels. Agarose hydrogels carrying covalently immobilized LN oligopeptides thus evoke specific responses from cells which contain receptors to the peptides used. Agarose hydrogels derivatized with neurite promoting peptide sequences may find applications in various models of in vivo regeneration of nervous tissue.

Biomimetic interfacial interpenetrating polymer networks control neural stem cell behavior

“…These results further confirm that the RGD peptide-modified IPN, without introducing any cooperative effects from mechanisms involving lam-IKVAV (19), was able to functionally substitute for laminin in early differentiation stages of adult NSCs. Furthermore, in contrast to fetal neuronal progenitors 14 or mature neurons, 12,13 IKVAV did not support the attachment, neuronal differentiation, or neurite extension of adult NSCs at the surface densities tested.…”

“…[11][12][13] We next tested whether another laminin motif, Ile-Lys-Val-Ala-Val [lam-IKVAV (19)], putatively involved in neurite extension of mature neurons 12,13 and differentiation of fetal neuronal progenitors, 14 could modulate cell behavior. We took advantage of the fact that the highly modular synthetic IPN matrix could be conjugated with diverse combinations of biochemical signals, such as lam-IKVAV(19) and bsp-RGD (15) peptides at various ratios (i.e., creation of binary peptide phase diagrams).…”

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

“…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 ).…”

Bioactive Nanomaterials for Neural Engineering

“…Collagen, fi bronectin, and laminin are examples of some major components of the ECM that have been used for this purpose (Yu and Bellamkonda 2003 ;Armstrong et al 2007 ;Koh et al 2010 ). Laminin, in particular, has been used most frequently for surface modifi cation or ECM mimicking purposes among others due to its ability to improve neurite extension and provide Schwann cell adhesion, proliferation, and migration (Yu and Bellamkonda 2003 ;Silva et al 2004 ;Yu et al 1999 ;Itoh et al 2001 ;Rangappa et al 2000 ;Rutkowski et al 2004 ;Toba et al 2001 ;Matsumoto et al 2000 ;Koh et al 2010 ;Bellamkonda et al 1995 ). Collagen and fi bronectin also have regenerative capacity in terms of Schwann cell adhesion, proliferation, and neurite outgrowth improvement; however, outcomes have shown to be signifi cantly lower than that of laminin in terms of regeneration (Yu and Bellamkonda 2003 ;Armstrong et al 2007 ;Koh et al 2010 ).…”