Enhanced neuronal differentiation in a three‐dimensional collagen‐hyaluronan matrix

Abstract

Efficient 3D cell systems for neuronal induction are needed for future use in tissue regeneration. In this study, we have characterized the ability of neural stem/progenitor cells (NS/PC) to survive, proliferate, and differentiate in a collagen type I-hyaluronan scaffold. Embryonic, postnatal, and adult NS/PC were seeded in the present 3D scaffold and cultured in medium containing epidermal growth factor and fibroblast growth factor-2, a condition that stimulates NS/PC proliferation. Progenitor cells from the embryonic brain had the highest proliferation rate, and adult cells the lowest, indicating a difference in mitogenic responsiveness. NS/PC from postnatal stages down-regulated nestin expression more rapidly than both embryonic and adult NS/PC, indicating a faster differentiation process. After 6 days of differentiation in the 3D scaffold, NS/PC from the postnatal brain had generated up to 70% neurons, compared with 14% in 2D. NS/PC from other ages gave rise to approximately the same proportion of neurons in 3D as in 2D (9-26% depending on the source for NS/PC). In the postnatal NS/PC cultures, the majority of betaIII-tubulin-positive cells expressed glutamate, gamma-aminobutyric acid, and synapsin I after 11 days of differentiation, indicating differentiation to mature neurons. Here we report that postnatal NS/PC survive, proliferate, and efficiently form synapsin I-positive neurons in a biocompatible hydrogel.

3D culture of adult mouse neural stem cells within functionalized self-assembling peptide scaffolds

“…17 Another study used a collagen and hyaluronan scaffold to grow embryonic, postnatal, and adult neural stem and progenitor cells. 18 Early differentiating human embryonic stem cells were further induced to differentiate in a 3D scaffold made of poly(lactic-co-glycolic acid)/poly(L-lactic acid). 19 Finally, one study compared the viability of hippocampal neurons and astrocytes grown on 2D conditions with that of cells grown on an aragonite 3D scaffold prepared from coralline exoskeleton, showing better results for the 3D cultures.…”

State-of-the-art of 3D cultures (organs-on-a-chip) in safety testing and pathophysiology

“…Functional synapses have been observed in three-dimensional neuronal cultures at similar or earlier time points compared to 2D models (O'Shaughnessy et al, 2003;van Vliet et al, 2007). Furthermore, neural stem/progenitor three-dimensional cell cultures showed an increased differentiation into neurons compared with 2D cultures (Brannvall et al, 2007). the phenotype of microglial cells is highly dependent on the culture conditions.…”

“…Moreover three-dimensional cultures often show enhanced survival and neuronal differentiation compared to traditional monolayer cultures (Brannvall et al, 2007;Peretz et al, 2007). the enhanced survival could be related to the improved cell-cell and cell-matrix contact, as it improves cell signaling and gap junction connections, but these factors may need further investigation, as also in vivo survival after, e.g., injection of cells in the brain is still not clarified (Hansson et al, 2000;Schierle et al, 1999).…”

Fluid Flow Shear Stress Stimulation on a Multiplex Microfluidic Device for Rat Bone Marrow Stromal Cell Differentiation Enhancement

“…As to the micro-environment cues, Akhavan et al reported accelerated differentiation of neural stem cells cultivated on ginseng-reduced graphene oxide sheets [33]. It is addressed that a three-dimensional collagen-hyaluronan matrix could enhance neuronal differentiation [34]. Francis et al demonstrated that preconditioning the human embryonic stem cell with hypoxic environment and its derived cytoprotective phenotype may promote the neural differentiation and enhanced cell survival rates [35].…”