Abstract
3D cell cultures are becoming more and more important in the field of regenerative medicine due to their ability to mimic the cellular physiological microenvironment. Among the different types of 3D scaffolds, we focus on the Nichoid, a miniaturized scaffold with a structure inspired by the natural staminal niche. The Nichoid can activate cellular responses simply by subjecting the cells to mechanical stimuli. This kind of influence results in different cellular morphology and organization, but the molecular bases of these changes remain largely unknown. Through RNA-Seq approach on murine neural precursors stem cells expanded inside the Nichoid, we investigated the deregulated genes and pathways showing that the Nichoid causes alteration in genes strongly connected to mechanobiological functions. Moreover, we fully dissected this mechanism highlighting how the changes start at a membrane level, with subsequent alterations in the cytoskeleton, signaling pathways, and metabolism, all leading to a final alteration in gene expression. The results shown here demonstrate that the Nichoid influences the biological and genetic response of stem cells thorough specific alterations of cellular signaling. The characterization of these pathways elucidates the role of mechanical manipulation on stem cells, with possible implications in regenerative medicine applications.
Highlights
Nowadays, three-dimensional (3D) scaffolds are deeply investigated for their ability to guide cell fate [1,2]
Among the different types of 3D scaffolds, we focus on the Nichoid, a miniaturized scaffold with a structure inspired by the natural staminal niche
Through RNA-Seq approach on murine neural precursors stem cells expanded inside the Nichoid, we investigated the deregulated genes and pathways showing that the Nichoid causes alteration in genes strongly connected to mechanobiological functions
Summary
Three-dimensional (3D) scaffolds are deeply investigated for their ability to guide cell fate [1,2]. Biocompatible scaffolds have been developed to mimic the physiological microenvironment of specific cells and their growth inside these scaffolds typically allows a cellular response more representative of the in vivo behavior with respect to the 2D cell culture conditions [3,4,5,6] For this reason, the use of 3D scaffolds is important in the field of regenerative medicine in order to improve stem cells therapeutic effect [7,8]. The “Nichoid,” a 3D scaffold inspired by the natural staminal niche, seems to have a great potential in the development of 3D cultures and could be of great relevance in the field of regenerative medicine [9,10,11] (Carelli et al 2020, accepted in Nanotheranostics). Recent analysis on the neural stem cells niche have highlighted an increased stiffness in this part of the brain [18]
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