Structural and functional characterization of a 3D neuroblastoma cell- based platform (in scaffold) for drug screening

Abstract

It is widely accepted that cells grown in three-dimension (3D) culture more accurately mimic in vivo microenvironment. However, with the wide range of physical, chemical and spatio properties required for 3D culture, the establishment of a three-dimensionality marker that can strike a balance between providing minimum environmental cues and showing Complex Physiological Relevance (CPR) is needed. Here CPR refers to structure and/or function mimicking exhibited in vivo and in 3D but not in 2D cultures. To validate the potential 3D biomarkers, the establishment of end-point CPR is desired. Among the four tissue types, neuronal cell type end-point CPR still remains elusive. This study showed that scaffold culture system (3D) enables cells to have morphologies closer to their in vivo counterpart in comparison to 2D cultures. Also, 3D culture has been shown to promote the differentiation of SH-SY5Y cells without any differentiation agent. It was demonstrated that the cytosolic calcium oscillation/spikes in 3D are statistically higher than that of 2D, which suggests calcium oscillation/spikes can serve as functional CPR for neuronal cell type. Additionally, it was shown that caveolae, the omega or cup-like invagination on cell membrane that are signaling channels enriched, density is higher in 3D than 2D. This finding may explain the calcium oscillation/spikes difference between 2D and 3D.