ABSTRACT
Human pluripotent stem cell-derived neural crest stem cells (NCSCs) provide a promising cell source for generating Schwann cells in the treatment of neurodegenerative diseases and traumatic injuries in the peripheral nervous system. Influencing cell behavior through a synthetic matrix topography has been shown to be an effective approach to directing stem cell proliferation and differentiation. Here we have investigated the effect of nanofiber topography on the differentiation of human embryonic stem cell-derived NCSCs towards the Schwann cell lineage. Using electrospun fibers of different diameters and alignments we demonstrated that aligned fiber matrices effectively induced cell alignment, and that fiber matrices with average diameters of 600nm and 1.6µm most effectively promoted NCSC differentiation towards the Schwann cell lineage compared with random fibers and two-dimensional tissue culture plates. More importantly, human NCSCs that were predifferentiated in Schwann cell medium for 2weeks exhibited higher sensitivity to the aligned fiber topography than undifferentiated NCSCs. This study provides an efficient protocol for Schwann cell derivation by combining an aligned nanofiber matrix and an optimized differentiation medium, and highlights the importance of matching extrinsic matrix signaling with cell intrinsic programming in a temporally specific manner.
Subject(s)
Extracellular Matrix/chemistry , Nanofibers/chemistry , Nanofibers/ultrastructure , Nanostructures/chemistry , Nanostructures/ultrastructure , Neural Crest/cytology , Neural Stem Cells/cytology , Schwann Cells/cytology , Biomimetic Materials/chemistry , Cell Differentiation , Cell Polarity , Cells, Cultured , Crystallization/methods , Electrochemistry/methods , Humans , Materials Testing , Particle SizeABSTRACT
Hyaluronic acid (HA) was added into fibroin solution to prepare fibroin-based porous composite scaffolds. HA exhibited important effects on pore formation and hydrophilicity of fibroin-based scaffold. The aqueous-fibroin/HA scaffolds had highly homogeneous and interconnected pores with porosity of above 90% and controllable pore size ranging from 123 to 253 microm. The water take-up of fibroin/HA scaffolds increased significantly with the increase of HA content. Containing HA at a defined content range, such as 3-6%, fibroin-based scaffolds' affinity to primary neural cells was improved. In 6%HA/fibroin scaffolds, neurosphere-forming cell migrated from their original aggregate and adhered tightly to the surface of scaffolds.
Subject(s)
Fibroins/chemistry , Fibroins/pharmacology , Hyaluronic Acid/chemistry , Hyaluronic Acid/pharmacology , Tissue Scaffolds/chemistry , Animals , Biomechanical Phenomena , Bombyx , Cell Adhesion/drug effects , Cell Proliferation/drug effects , Female , Fibroins/metabolism , Hyaluronic Acid/metabolism , Hydrophobic and Hydrophilic Interactions , Insect Proteins/chemistry , Insect Proteins/metabolism , Insect Proteins/pharmacology , Microscopy, Electron, Scanning , Neurons/cytology , Neurons/drug effects , Porosity/drug effects , Pregnancy , Rats , Rats, Sprague-Dawley , Spectroscopy, Fourier Transform Infrared , Stem Cells/cytology , Stem Cells/drug effects , Tissue Engineering , Water/chemistryABSTRACT
Neural stem cells (NSCs) cultured on glass surfaces modified by different chemical groups, including hydroxyl (-OH), sulfonic (-SO3H), amino (-NH2), carboxyl (-COOH), mercapto (-SH) and methyl (-CH3) groups, are shown here to commit to phonotypes with extreme sensitivity to surface chemical groups. The adhering NSCs at the level of single cells exhibited morphological changes in response to different chemical groups. NSCs on -SO(3)H surfaces had the largest contact area and the most flattened morphology, while those on -CH(3) surfaces exhibited the smallest contact area and the most rounded morphology. After 5 days of culture, the migration of NSCs from their original aggregates onto these test surfaces followed the trend: -NH2>-COOH=-SH>>-SO3H>-CH3>-OH. The expression of specific markers, including nestin, beta-Tubulin-III, glial fibrillary acidic protein and O4, were used to examine NSCs lineage specification. The -SO3H surfaces favored NSCs differentiation into oligodendrocytes, while NSCs in contact with -COOH, -NH2, -SH and -CH3 had the ability to differentiate into neurons, astrocytes and oligodendrocytes. Compared to -COOH surfaces, -NH2 seemed to promote neuronal differentiation. These chemically modified surfaces exhibited regulation of NSCs on adhesion, migration and differentiation potential, providing chemical means for the design of biomaterials to direct NSCs lineage specification for neural tissue engineering.
