3D Culturing and differentiation of SH-SY5Y neuroblastoma cells on bacterial nanocellulose scaffolds

• Published in: Artificial cells, nanomedicine, and biotechnology Vol. 42; no. 5; pp. 302 - 308
• Main Authors: Innala, Marcus, Riebe, Ilse, Kuzmenko, Volodymyr, Sundberg, Johan, Gatenholm, Paul, Hanse, Eric, Johannesson, Sara
• Format: Journal Article
• Language: English
• Published: England Informa Healthcare 01.10.2014; Taylor & Francis
• Subjects: bacterial nanocellulose; Basic Medicine; Cell Culture Techniques - methods; Cell Differentiation - drug effects; Cell Line, Tumor; Cellulose - chemistry; Cellulose - metabolism; Cellulose - pharmacology; Gluconacetobacter xylinus - metabolism; Humans; Medicinska grundvetenskaper; Nanostructures - chemistry; Neuroblastoma - pathology; neuronal network 3D model; scaffolds; SH-SY5Y cells; surface modification; scaffolds; SH-SY5Y cells; surface modification; bacterial nanocellulose; neuronal network 3D model
• ISSN: 2169-1401; 2169-141X; 2169-141X
• DOI: 10.3109/21691401.2013.821410

Abstract A new in vitro model, mimicking the complexity of nerve tissue, was developed based on a bacterial nanocellulose (BNC) scaffold that supports 3D culturing of neuronal cells. BNC is extracellularly excreted by Gluconacetobacter xylinus (G. xylinus) in the shape of long non-aggregated nanofibrils. The cellulose network created by G. xylinus has good mechanical properties, 99% water content, and the ability to be shaped into 3D structures by culturing in different molds. Surface modification with trimethyl ammonium beta-hydroxypropyl (TMAHP) to induce a positive surface charge, followed by collagen I coating, has been used to improve cell adhesion, growth, and differentiation on the scaffold. In the present study, we used SH-SY5Y neuroblastoma cells as a neuronal model. These cells attached and proliferated well on the BNC scaffold, as demonstrated by scanning electron microscopy (SEM) and the 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium) (MTS) assay. Following neuronal differentiation, we demonstrated functional action potentials (APs) by electrophysiological recordings, indicating the presence of mature neurons on the scaffolds. In conclusion, we have demonstrated for the first time that neurons can attach, proliferate, and differentiate on BNC. This 3D model based on BNC scaffolds could possibly be used for developing in vitro disease models, when combined with human induced pluripotent stem (iPS) cells (derived from diseased patients) for detailed investigations of neurodegenerative disease mechanisms and in the search for new therapeutics.