Experimental Cell Transplantation Group, Laboratory of Experimental Hematology, University of Antwerp, Antwerp, Belgium; Vaccine and Infectious Disease Institute (Vaxinfectio), University of Antwerp, Antwerp, Belgium.
Vaccine and Infectious Disease Institute (Vaxinfectio), University of Antwerp, Antwerp, Belgium.
Biomaterials. 2015 Feb;41:122-31. doi: 10.1016/j.biomaterials.2014.11.025. Epub 2014 Dec 5.
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.
神经干细胞（NSC）在病变或损伤脑组织中的移植已被广泛研究，作为治疗各种神经病理学的一种潜在方法。然而，有效的细胞替代治疗依赖于细胞移植物在移植后克服缺氧和/或免疫屏障的内在能力。在这种情况下，假设对移植的 NSC 进行结构支持将是至关重要的。在这项研究中，我们提出了一种新的用于 1.5 毫米厚的小鼠脑切片的去细胞化方案，从而产生了无细胞的三维（3D）脑切片。接下来，将获得的 3D 脑切片接种表达绿色荧光蛋白和荧光素酶报告蛋白的小鼠 NSC（NSC-eGFP/Luc）。使用实时生物发光成像，在培养中对接种的 NSC-eGFP/Luc 细胞的存活和生长进行了 1-7 周的纵向监测，表明无细胞脑切片能够支持 NSC 的持续体外生长。接下来，使用共聚焦显微镜检查了 3D 迷宫样细胞结构的组织。此外，在有丝分裂刺激（EGF 和 hFGF-2）下，这种 3D 培养中的大多数细胞保留了它们的 NSC 表型。总之，我们在这里提出了一种新的小鼠脑切片去细胞化方案，该方案随后支持未分化 NSC 的长期 3D 培养。
