3D culture of murine neural stem cells on decellularized mouse brain sections

• Published in: Biomaterials Vol. 41; pp. 122 - 131
• Main Authors: De Waele, Jorrit, Reekmans, Kristien, Daans, Jasmijn, Goossens, Herman, Berneman, Zwi, Ponsaerts, Peter
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier Ltd 01.02.2015
• Subjects: 3D cell culture; Advanced Basic Science; Animals; Bioluminescence imaging; Brain; Brain - cytology; Cell Culture Techniques - methods; Cell Proliferation; Cell Survival; Cells, Cultured; Cellular; Culture; Decellularization; Dentistry; Genes, Reporter; Green Fluorescent Proteins - metabolism; Growth scaffold; Imaging; Luciferases - metabolism; Medical services; Mice, Inbred C57BL; Neural stem cells; Neural Stem Cells - cytology; Neural Stem Cells - metabolism; Phenotype; Stem cells; Stimuli; Three dimensional; Transplantation; Brain; Neural stem cells; 3D cell culture; Bioluminescence imaging; Growth scaffold; Decellularization
• ISSN: 0142-9612; 1878-5905
• DOI: 10.1016/j.biomaterials.2014.11.025

Abstract Transplantation of neural stem cells (NSC) in diseased or injured brain tissue is widely studied as a potential treatment for various neurological pathologies. However, effective cell replacement therapy relies on the intrinsic capacity of cellular grafts to overcome hypoxic and/or immunological barriers after transplantation. In this context, it is hypothesized that structural support for grafted NSC will be of utmost importance. With this study, we present a novel decellularization protocol for 1.5 mm thick mouse brain sections, resulting in the generation of acellular three-dimensional (3D) brain sections. Next, the obtained 3D brain sections were seeded with murine NSC expressing both the eGFP and luciferase reporter proteins (NSC-eGFP/Luc). Using real-time bioluminescence imaging, the survival and growth of seeded NSC-eGFP/Luc cells was longitudinally monitored for 1–7 weeks in culture, indicating the ability of the acellular brain sections to support sustained ex vivo growth of NSC. Next, the organization of a 3D maze-like cellular structure was examined using confocal microscopy. Moreover, under mitogenic stimuli (EGF and hFGF-2), most cells in this 3D culture retained their NSC phenotype. Concluding, we here present a novel protocol for decellularization of mouse brain sections, which subsequently support long-term 3D culture of undifferentiated NSC.