In Vitro 3D Regeneration ‐like Growth of Human Patient Brain Tissue

Abstract In vitro culture of primary neurons are widely adapted with embryonic but not mature brain tissue. Here, we extended a previously developed bioengineered 3D embryonic brain tissue model to resected normal patient brain tissue in an attempt to regenerate human neurons in vitro. Single cells and small sized (diameter < 100 μm) spheroids from dissociated brain tissue were seeded into 3D silk fibroin‐based scaffolds, with or without collagen or Matrigel, and compared with 2D cultures and scaffold‐free suspension cultures. Changes of cell phenotypes (neuronal, astroglial, neural progenitor and neuroepithelial) were quantified with flow cytometry and analyzed with a new method of statistical analysis specifically designed for percentage comparison. Compared to a complete lack of viable cells in conventional neuronal cell culture condition, supplements of VEGF‐containing pro‐endothelial cell condition led to regenerative growth of neurons and astroglial cells from ‘normal' human brain tissue of epilepsy surgical patients. This process involved delayed expansion of Nestin+ neural progenitor cells, emergence of TUJ1+ immature neurons, and Vimentin+ neuroepithelium‐like cell sheet formation in prolonged culture (14wks). Micro‐tissue spheroids, but not single cells, supported the brain tissue growth, suggesting importance of preserving native cell‐cell interactions. The presence of 3D scaffold, but not hydrogel, allowed for Vimentin+ cell expansion, indicati...
Source: Journal of Tissue Engineering and Regenerative Medicine - Category: Biotechnology Authors: Tags: RESEARCH ARTICLE Source Type: research