Exploration of physical and chemical cues on retinal cell fate

Abstract

Identification of the key components in the physical and chemical milieu directing donor cells into a desired phenotype is a requirement in the investigation of bioscaffolds for the advancement of cell-based therapies for retinal neurodegeneration. We explore the effect of electrospun poly-ε-caprolactone (PCL) fiber scaffold topography and functionalization and culture medium, on the behavior of mouse retinal cells. Dissociated mouse retinal post-natal cells were seeded on random or aligned oriented fibers, with or without laminin coating and cultured with either basic or neurotrophins enriched medium for 7days. Addition of laminin in combination with neurotrophins clearly promoted cell- morphology, fate, and neurite extension. Nanotopography per se significantly affected cell morphology, with mainly bipolar profiles on aligned fibers and more multipolar profiles on random fibers. Laminin induced a remarkable 90° switch of neurite orientation. Herewith, we demonstrate that the chemical cue is stronger than the physical cue for the orientation of retinal neurites and describe the requirement of both neurotrophins and extracellular matrix proteins for extended neurite outgrowth and formation of complex retinal neuronal networks. Therefore, tailor-made PCL fiber mats, which can be physically and chemically modified, indeed influence cell behavior and hence motivate further retinal restorative studies using this system.

Immunocytochemical Profiling of Cultured Mouse Primary Retinal Cells

“…13 Retinal cells when dissociated suffer mechanical stress such as breakage of their neurites and axon. Central nervous system neurons, including retinal neurons, lack regeneration capabilities but retain their neuritogenic potential under certain conditions in vivo and in vitro.…”

“…This medium was formulated to culture retinal ganglion cells (RGCs) after immunopanning 20,25 and also recently used by us for the efficient culture of the major retinal cell types at fibrous substrates. 13 …”

“…6–10 Lately, crucial chemical and physical factors (including bioscaffolds) for optimal donor cell development and administration have been studied by others and us. 11–13 …”

Tailor-Made Electrospun Culture Scaffolds Control Human Neural Progenitor Cell Behavior—Studies on Cellular Migration and Phenotypic Differentiation

“…Although promising, increased understanding on the key physicaland chemical cues of electrospun fibrous substrates for supporting survival-and controlling neural cell behavior is needed for further advancements in many research fields in neuroscience. Indeed, recently, for the first time we reported a significant effect of chemical respective physical cues on post-natal mouse retinal cell behavior using different designed electrospun fibrous scaffolds [24]. Nanotopography per se significantly affected cell morphology, but the chemical cue, i.e.…”

Author Correction: Electrospun nerve guide conduits have the potential to bridge peripheral nerve injuries in vivo

“…“PLC nerve guide characteristics. (A) SEM images of the PCL guide with more defined number of micro channels; (B) individual channel dimensions traced in ImageJ to calculate the porosity; (C) randomly oriented PCL fibers 1 ; and (D) aligned PCL fibers 1 , indicating the porosity of the scaffold, and (E) the Fourier spectra of the random and aligned fibers. (F) SEM image of the PLLA nerve guide, where the pores appear to be less well defined than in the PCL guide; (G) the PCL nerve guide inserted in its silicone shell; (H) explanted nerve guide integrated in the regenerating sciatic nerve; (I) hollow tube bridged by a thin silicone matrix; (J) the regenerated matrix measured to the length of the sciatic nerve defect of 10 mm”.…”