| An in vitro Model of Neuron-macrophage Interaction to Generate Macrophages with Neurite Outgrowth Properties
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Author:
Date:
2016-11-20
[Abstract] Macrophages are known to play beneficial roles in axon regeneration after nerve injury. To develop an in vitro model in which injury signals can elicit pro-regenerative macrophage activation, we established co-cultures consisting of adult dorsal root ganglia sensory neurons and peritoneal macrophages and added cAMP analogue dibutyryl cAMP. The conditioned medium collected from the co-cultures exhibited robust neurite outgrowth activities. The neurite outgrowth activities were almost completely abrogated by addition of minocycline, a macrophage deactivator, indicating that factors responsible for neurite outgrowth are produced by activated macrophages.
[摘要] 已知巨噬细胞在神经损伤后的轴突再生中发挥有益作用。 为了开发一种体外模型，其中损伤信号可以引发再生巨噬细胞活化，我们建立了由成体背根神经节感觉神经元和腹膜巨噬细胞组成的共培养物，并加入cAMP类似物二丁酰基cAMP。 从共同培养物收集的条件培养基表现出强烈的神经突生长活动。 通过添加米诺环素（巨噬细胞减活剂）几乎完全消除神经突生长活动，表明负责神经突生长的因子是由活化的巨噬细胞产生的。
【背景】成年哺乳动物的CNS神经元在损伤后不会自发再生轴突。 预处理周围神经损伤允许背根神经节（DRG）感觉轴突通过促进再生相关基因的表达来再生中心分支。 我们以前已经表明，预处理损伤后DRG中的活化巨噬细胞有助于提高DRG感觉神经元的内在再生能力（Kwon等，2013）。 为了确定参与神经损伤后巨噬细胞活化的分子因子，我们开发了体外模型，其中神经元 - 巨噬细胞相互作用由cAMP引起，cAMP是增强神经元再生能力的众所周知的试剂。 与使用酵母聚糖激活巨噬细胞的以前的模型相比，我们的模型在预处理外周损伤模型中使用类似于分子事件的更多的生理刺激。
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| Establishing Primary Malignant Pleural Mesothelioma (MPM) Cell Cultures
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Author:
Date:
2012-11-05
[Abstract] This is a general protocol for the isolation and maintenance of primary MPM cultures as a tool for the identification of Tumor Initiating Cells and early progenitor-targeting drugs (Cioce et al., 2010). Primary cultures can be propagated efficiently for 8-12 weeks and xenotransplanted in NOD/SCID mice while retaining the histofeatures of the originating tumor (Canino et al., 2012). The protocol is suitable for both MPM solid specimens and pleural effusion. For increased clarity, initially two separate sections addressing the isolation of MPM cells from solid tumors and pleural effusions are here provided.
[摘要] 这是用于分离和维持初级MPM培养物的一般方案，作为鉴定肿瘤启动细胞和早期祖细胞靶向药物的工具（Cioce等人，2010）。 原代培养物可以有效繁殖8-12周，并且在NOD/SCID小鼠中异种移植，同时保留起源肿瘤的组织特征（Canino等人，2012）。 该方案适用于MPM固体标本和胸腔积液。 为了增加清晰度，最初提供了两个分开的部分，其涉及从实体瘤和胸腔积液中分离MPM细胞。
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