Lee Jea-Young, Lin Roger, Nguyen Hung, Grant Liska M, Lippert Trenton, Kaneko Yuji, Borlongan Cesar V
Department of Neurosurgery and Brain Repair, Center of Excellence for Aging and Brain Repair, University of South Florida Morsani College of Medicine, Tampa, FL, USA.
Center of Excellence for Aging & Brain Repair, Morsani College of Medicine, University of South Florida, 12901 Bruce B. Downs Blvd., MDC 78, Tampa, FL, 33612, USA.
Methods Mol Biol. 2019;2045:299-310. doi: 10.1007/7651_2018_121.
Traumatic brain injury (TBI) displays cognitive and motor symptoms following the initial injury which can be exacerbated by secondary cell death. Aging contributes significantly to the morbidity of TBI, with higher rates of negative neurological and behaviors outcomes. In the recent study, young and aged animals were injected intravenously with human adipose-derived mesenchymal stem cells (hADSCs) (Tx), conditioned media (CM), or vehicle (unconditioned media) following TBI. The beneficial effects of hADSCs were analyzed using various molecular and behavioral techniques. More specially, DiR-labeled hADSCs were used to observe the biodistribution of the transplanted cells. In addition, a battery of behavior tests was conducted to evaluate the neuromotor function for each treatment group and various regions of the brain were analyzed utilizing Nissl, hematoxylin and eosin (H&E), and human nuclei (HuNu) staining. Finally, flow cytometry was also performed to determine the levels of various proteins in the spleen. Here, we discuss the protocols for characterizing the histopathological and behavioral effects of transplanted stem cells in an animal model of TBI, with an emphasis on the role of aging in the therapeutic outcomes.
创伤性脑损伤（TBI）在初始损伤后会出现认知和运动症状，继发性细胞死亡会使这些症状加重。衰老对TBI的发病率有显著影响，神经和行为不良后果的发生率更高。在最近的研究中，年轻和老年动物在TBI后静脉注射人脂肪间充质干细胞（hADSCs）（Tx）、条件培养基（CM）或载体（未处理培养基）。使用各种分子和行为技术分析了hADSCs的有益效果。更具体地说，使用DiR标记的hADSCs来观察移植细胞的生物分布。此外，进行了一系列行为测试以评估每个治疗组的神经运动功能，并利用尼氏染色、苏木精和伊红（H&E）染色以及人类细胞核（HuNu）染色对大脑的各个区域进行分析。最后，还进行了流式细胞术以确定脾脏中各种蛋白质的水平。在此，我们讨论在TBI动物模型中表征移植干细胞的组织病理学和行为效应的方案，重点是衰老在治疗结果中的作用。
