Genomic and Morphological Changes of Neuroblastoma Cells in Response to Three-Dimensional Matrices

Abstract

Advances in neural tissue engineering require a comprehensive understanding of neuronal growth in 3 dimensions. This study compared the gene expression of SH-SY5Y human neuroblastoma cells cultured in 3-dimensional (3D) with those cultured in 2-dimensional (2D) environments. Microarray analysis demonstrated that, in response to varying matrix geometry, SH-SY5Y cells exhibited differential expression of 1,766 genes in collagen I, including those relevant to cytoskeleton, extracellular matrix, and neurite outgrowth. Cells extended longer neurites in 3D collagen I cultures than in 2D. Real-time reverse transcriptase polymerase chain reaction experiments and morphological analysis comparing collagen I and Matrigel tested whether the differential growth and gene expression reflected influences of culture dimension or culture material. SH-SY5Y neuroblastoma cells responded to geometry by differentially regulating cell spreading and genes associated with actin in similar patterns for both materials; however, neurite outgrowth and the expression of the gene encoding for neurofilament varied with the type of material. Electron microscopy and mechanical analysis showed that collagen I was more fibrillar than Matrigel, with larger inter-fiber distance and higher stiffness. Taken together, these results suggest complex cell-material interactions, in which the dimension of the culture material influences gene expression and cell spreading and the structural and mechanical properties of the culture material influence gene expression and neurite outgrowth.

The design of electrospun PLLA nanofiber scaffolds compatible with serum-free growth of primary motor and sensory neurons

“…Instead of using primary neurons harvested from the nervous system, most in vitro testing of biomaterials relies on the use of renewable cell lines [1,2,[9][10][11][12][13] or, more rarely, primary cells grown in media supplemented with serum [1,3,14,15]. Both of these approaches have significant drawbacks.…”

Three-Dimensional Neural Spheroid Culture: AnIn VitroModel for Cortical Studies

“…1 Microenvironmental cues, such as cellular, biochemical, mechanical, and topographical cues, can influence cell behaviors, including viability, proliferation, differentiation, migration, and protein and gene expressions. [2][3][4][5][6] Notably, many native microenvironmental cues are lost in traditional twodimensional (2D) cultures, in which cells spread on substrates that are often several orders of magnitudes stiffer than brain tissue and lack the brain's extracellular matrix (ECM) organization. 7 Cells in 2D are forced to adapt a planar morphology and can form intercellular connections only in the lateral direction.…”

“…1,8 For example, 3D culture of human induced pluripotent stem cells has enabled the study of microcephaly, 9 and the hallmarks of Alzheimer's disease, amyloid-b plaques, and neurofibrillary tangles, have been more effectively modeled in 3D cultures than in 2D. 10 Historically, 3D neural cultures were developed using scaffolds, 3,[11][12][13][14] and complex cell compartmentalization and organization have been achieved through scaffold design. [15][16][17] Recently, neural cells have been cultured through self-assembly into spheroid structures in which they can produce their own matrix; 18,19 these spheroids have been suggested to approximate in vivo-like cell behavior better than scaffold-based cultures.…”

Estudo das células Neuro2A sobre os biomateriais PCL e PLLA

“…Sabe-se que a adesão, migração, diferenciação e a proliferação celular sobre determinado substrato depende em parte das propriedades intrínsecas de cada tipo celular, bem como, o tipo de material utilizado [1][2][3][4] . Dentre os produtos celulares, pode-se ressaltar a síntese de moléculas de adesão celular, como laminina, fibronectina e colágeno, importantes para o cultivo e o crescimento celular [5,6] . Além disso, alguns experimentos in vitro demonstraram que a utilização de fatores de crescimento e outras substâncias como o interferon e acetato de glatiramer provocam a proliferação celular [7,8] .…”