3D bioprinting of reactive astrocyte-derived neural stem cells

Abstract

In vitro models for the study of physiological processes and disease development of the Central Nervous System (CNS) have several limitations, such as poor representation of the three-dimensional (3D) cellular environment. As an alternative to two-dimensional (2D) cell cultures, 3D bioprinting represents an alternative to produce artificial, tissue-like structures with the possibility of modulating the microenvironment. Several cell types can be used in 3D bioprinting, and stem cells are ideal candidates for this purpose. Neural stem cells are found primarily in the canonical neurogenic niches of the CNS, thus characterizing the limited neurogenic potential of nervous tissue. Recently, an alternative for obtaining neural stem cells from reactive astrocytes has been described and the 3D bioprinting of these cells is unprecedented. Astrocytes make up a heterogeneous population with different capacities to generate stem cells after activation. The analysis of differentiation and proliferation, taking into account the population heterogeneity of stem cells derived from reactive bioprinted astrocytes, may validate the use of this model to reproduce physiological processes of the CNS. Thus, we propose to evaluate the cultivation of neural stem cells derived from 3D bioprinted astrocytes, focusing on the analysis of global transcriptomics by Next Generation Sequencing (NGS). In addition, the quantitative and qualitative analysis of differentiation and proliferation by qPCR and immunofluorescence will be carried, comparing 3D bioprinted cells with those kept in a 2D environment and from an in vivo model. (AU)