3D culture of murine neural stem cells on decellularized mouse brain sections
3D culture of murine neural stem cells on decellularized mouse brain sections
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DOI：
10.1016/j.biomaterials.2014.11.025
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发表时间：
2015-02-01
期刊：
影响因子：
14
通讯作者：
Ponsaerts, Peter
中科院分区：
文献类型：
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作者：
De Waele, Jorrit;Reekmans, Kristien;Ponsaerts, Peter
神经干细胞在病变或损伤脑组织中的移植作为一种潜在的治疗方法被广泛研究。然而，有效的细胞替代疗法依赖于细胞移植物在移植后克服缺氧和/或免疫障碍的内在能力。在这种情况下，假设移植NSC的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1.5 mm厚小鼠脑切片脱细胞方案，从而产生无细胞三维（3D）脑切片。接下来，将获得的3D脑切片植入表达eGFP和荧光素酶报告蛋白（NSC-eGFP/Luc）的小鼠NSC。利用实时生物发光成像技术，在培养1-7周的时间内，对NSC- egfp /Luc种子细胞的存活和生长情况进行了纵向监测，表明脱细胞脑切片能够支持NSC的持续离体生长。接下来，使用共聚焦显微镜检查三维迷宫状细胞结构的组织。此外，在有丝分裂刺激（EGF和hFGF-2）下，3D培养中的大多数细胞保留了NSC表型。最后，我们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(C) 2014 Elsevier Ltd.)。版权所有。
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 (C) 2014 Elsevier Ltd. All rights reserved.