Functional maturation of human neural stem cells in a 3D bioengineered brain model enriched with fetal brain-derived matrix

Abstract

15Brain extracellular matrix (ECM) is often overlooked in vitro brain tissue models, despite its 16 instructive roles during development. Using developmental stage-sourced brain ECM in reproducible 3D 17 bioengineered culture systems, we demonstrate enhanced functional differentiation of human induced 18 neural stem cells (hiNSCs) into healthy neurons and astrocytes. Particularly, fetal brain tissue-derived 19 ECM supported long-term maintenance of differentiated neurons, demonstrated by morphology, gene 20 expression and secretome profiling. Astrocytes were evident within the second month of differentiation, 21 and reactive astrogliosis was inhibited in brain ECM-enriched cultures when compared to unsupplemented 22cultures. Functional maturation of the differentiated hiNSCs within fetal ECM-enriched cultures was 23 confirmed by calcium signaling and unsupervised cluster analysis. Additionally, the study identified 24 native biochemical cues in decellularized ECM with notable comparisons between fetal and adult brain-25 derived ECMs. The development of novel brain-specific biomaterials for generating mature in vitro brain 26 models provides an important path forward for interrogation of neuron-glia interactions. 27

Gene Expression of Mouse Hippocampal Stem Cells Grown in a Galactose-Derived Molecular Gel Compared to In Vivo and Neurospheres

“…Hence the scaffold cannot be considered just as a neutral support for the cell growth in 3D. Highlighting the impact of the scaffold on the gene expression of the cultivated cells is thus very important in order to better select the right scaffold for a given application [9,10]. Many biomaterials of different types are currently being developed for addressing in vitro 3D cell cultures of neuronal cells.…”

“…Many biomaterials of different types are currently being developed for addressing in vitro 3D cell cultures of neuronal cells. They are most often derived from the extracellular matrix (ECM) or its components [9][10][11] or made of bio sourced polymers [9,[12][13][14] and purely synthetic [15] or bio sourced/synthetic composites [11,16]. Emerging alternative biomaterials for neuron cell culture are selfassembling fibrillar scaffolds [17][18][19][20][21][22][23][24][25][26].…”