Neural differentiation directed by self‐assembling peptide scaffolds presenting laminin‐derived epitopes

Abstract

We prepared biofunctionalized matrices for cell growth using (RADA)(3)IKVAV(RADA)(3) ((Arg-Ala-Asp-Ala)(3)-Ile-Lys-Val-Ala-Val-(Arg-Ala-Asp-Ala)(3)) and (RADA)(4)IKVAV ((Arg-Ala-Asp-Ala)(4)-Ile-Lys-Val-Ala-Val), self-assembling peptides with a laminin-derived sequence inserted between and attached terminally to the repeats of RADA, respectively. The material-cell interactions were investigated with PC12, a cell line commonly used as a model for studying neural differentiation. The behavior of PC12 and especially the neural differentiation was guided by the presence of IKVAV. Furthermore, the cell-material interactions were dependent on the culture dimensionality and the position of IKVAV in the self-assembling peptide template. In the two-dimensional (2-D) culture, matrices containing IKVAV stimulated significantly longer neurite outgrowths from PC12 cells than did (RADA)(4). More pronounced effect was observed in (RADA)(3)IKVAV(RADA)(3) than in (RADA)(4)IKVAV. In the three-dimensional (3-D) culture, neurite outgrowth was not observed in the biofunctionalized matrices. Instead, cells displayed higher proliferation rate and survived longer culture time than in the 2-D culture, with such enhancement being most significant in (RADA)(3)IKVAV(RADA)(3.) Despite the lack of differentiation phenotype, the cells grown in 3-D biofunctionalized matrices were primed for differentiation, as evident by enhanced neurite outgrowth, increased neurite networking, and up-regulated expression of differentiation markers upon their reintroduction to the 2-D culture condition on petri dish. With the ease of incorporating biofunctional epitopes, and the flexibility to support either 2-D or 3-D culture, self-assembling peptides provide versatile scaffolds to study the multiple facets of biomaterial-cell interactions.

Differentiation Ability of Amniotic Fluid-Derived Stem Cells Cultured on Extracellular Matrix-immobilized Surface

“…Laminin, a molecule that is present in the basal lamina from the earliest stage of neural tube formation, has been reported to promote the proliferation and differentiation of neuroepithelial cells [46] and to induce neurite outgrowth in human mesenchymal stem cells [47,48]. However, no significant increase in the neural differentiation ratios of amniotic fluid stem cells cultured on lamininimmobilized (PS-LN-C and PS-LN-G) dishes was observed in this study.…”

Differentiation of Human Neural Progenitor Cells in Functionalized Hydrogel Matrices

“…19–21 Laminin-derived sequences have been shown to be supportive of neuronal differentiation and survival when inserted into scaffolds. 22 Functionalization of PuraMatrix with different short peptides, including among others a laminin-derived sequence (-GGSDPGYIGSR-) and a sequence found in the bone marrow homing factor (-GGPFSSTKT-), was shown to affect the mechanical properties of the matrix and alter neuronal differentiation. 15,16,23 In a recent study, we supplemented PuraMatrix with laminin and described the fate of human neural progenitor cells (hNPCs) encapsulated in these 3D scaffolds, regarding differentiation and survival.…”

Exploiting bacterial peptide display technology to engineer biomaterials for neural stem cell culture

“…[14] In addition the peptides were displayed on self-assembled monolayer surfaces that lose stability over time and contain surface defects where cells can attach non-specifically. [62–66] Yeast peptide display libraries have been used to target the α v β 3 integrin with RGD peptides,[24] but a method that is unbiased towards any particular motif is desirable, since it can allow the investigation of peptides for non-RGD binding cellular receptors.…”