Chiavari Marta, Ciotti Gabriella Maria Pia, Navarra Pierluigi, Lisi Lucia
Institute of Pharmacology, Catholic University Medical School, L.go F Vito 1, 00168 Rome, Italy.
Fondazione Policlinico Universitario Agostino Gemelli, L.go F. Vito 1, 00168 Rome, Italy.
Brain Sci. 2019 May 15;9(5):111. doi: 10.3390/brainsci9050111.
The characterization of human microglia has been hampered by poor availability of human cell sources. However, microglia is involved in the physiopathology of multiple sclerosis, Alzheimer's disease, Parkinson's disease, HIV dementia, retinal degenerative diseases, cancer, and many other conditions. Therefore, there is an important need to have experimental paradigms of human microglia characterized and usable to study the role of microglia in the different pathologies in which it is involved. In the present work, we carried out an extensive characterization of Immortalized Human Microglia-SV40 cell line (IMhu), marketed by Applied Biological Material. The functional response of IMhu to a large variety of stimuli was studied. In particular, we investigated morphology, mortality, and changes in the production of different cytokines and chemokines, both under basal conditions and after stimulation. Moreover, western blotting analysis was conducted on phospho-mTOR (Ser 2448) and downstream parameters, p-P70S6K and 4EBP1, in order to understand if IMhu can be used for evaluations of mTOR pathway. In conclusion, IMhu cells proved to be a useful experimental model to investigate the physiopathology of inflammatory disease that involved microglia cells, including pathological conditions that involved the mTOR pathway.
人类细胞来源的匮乏阻碍了对人类小胶质细胞的特性研究。然而，小胶质细胞参与了多发性硬化症、阿尔茨海默病、帕金森病、HIV 痴呆症、视网膜退行性疾病、癌症以及许多其他病症的病理生理过程。因此，迫切需要建立已表征的、可用于研究小胶质细胞在其所涉及的不同病理过程中作用的人类小胶质细胞实验范式。在本研究中，我们对应用生物材料公司销售的永生化人类小胶质细胞 - SV40 细胞系（IMhu）进行了广泛的特性研究。研究了 IMhu 对多种刺激的功能反应。具体而言，我们研究了基础条件下以及刺激后细胞的形态、死亡率以及不同细胞因子和趋化因子产生的变化。此外，对磷酸化 mTOR（Ser 2448）及其下游参数 p - P70S6K 和 4EBP1 进行了蛋白质印迹分析，以了解 IMhu 是否可用于评估 mTOR 信号通路。总之，IMhu 细胞被证明是一种有用的实验模型，可用于研究涉及小胶质细胞的炎症性疾病的病理生理过程，包括涉及 mTOR 信号通路的病理状况。
