Immunohistochemical Identification of Human Skeletal Muscle Macrophages
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Author:
Kate Kosmac, Bailey D. Peck, R. Grace Walton, Jyothi Mula, Philip A. Kern, Marcas M. Bamman, Richard A. Dennis, Cale A. Jacobs, Christian Lattermann, Darren L. Johnson and Charlotte A. Peterson,
Date:
2018-06-20
[Abstract] Macrophages have well-characterized roles in skeletal muscle repair and regeneration. Relatively little is known regarding the role of resident macrophages in skeletal muscle homeostasis, extracellular matrix remodeling, growth, metabolism and adaptation to various stimuli including exercise and training. Despite speculation into macrophage contributions during these processes, studies characterizing macrophages in non-injured muscle are limited and methods used to identify macrophages vary. A standardized method for the identification of human resident skeletal muscle macrophages will aide ...
[摘要] 巨噬细胞在骨骼肌修复和再生中具有很好的特征。关于驻留巨噬细胞在骨骼肌动态平衡,细胞外基质重塑,生长,代谢和适应各种刺激(包括运动和训练)中的作用知之甚少。尽管在这些过程中推测了巨噬细胞的贡献,但表征非受伤肌肉中的巨噬细胞的研究是有限的,用于鉴定巨噬细胞的方法各不相同。用于鉴定人类骨骼肌巨噬细胞的标准化方法将有助于鉴定这些免疫细胞,并可用于各研究结果的比较。在这里,我们提出免疫组织化学(IHC)协议,通过流式细胞术验证,以清楚地识别常驻人类骨骼肌巨噬细胞种群。我们显示CD11b和CD206双IHC有效识别人骨骼肌中的巨噬细胞。此外,非受伤人骨骼肌中的大多数巨噬细胞表现出“混合”M1 / M2表型,表达CD11b,CD14,CD68,CD86和CD206。在休息的骨骼肌中存在相对较少的CD11b + / CD206-巨噬细胞群。这种人口的相对丰度的变化可能反映了骨骼肌肉环境的重要变化。肌肉中的CD11b和CD206 IHC也显示出巨噬细胞的不同形态学特征,这些特征可能与这些细胞的功能状态有关。
【背景】巨噬细胞是能够适应局部微环境变化的多向性免疫细胞。在过去几年中,研究表明巨噬细胞表型是动态的,存在于连续统一体中(Mosser and Edwards,2008,Italiani and Boraschi,2014,Martinez and ...
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Measuring Rat Serum Osmolality by Freezing Point Osmometry
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Author:
Date:
2016-10-05
[Abstract] Blood serum or plasma osmolality is the measure of electrolyte to water balance in the body’s circulation, and is tightly regulated in physiological states in order to maintain normal levels of serum solute (Bourque, 2008). Osmolality is defined as the number of osmoles of solute per kg of water (mOsm/kg) (Dufour, 1993) and can be measured using different techniques that rely on the colligative properties of the solution. The most commonly used in lab settings are vapour pressure and freezing point osmometry, which are relatively quick and easy to perform. Freezing point osmometry is ...
[摘要] 血清或血浆渗透压是体内循环中电解质与水平衡的量度,并且在生理状态下受到严格的调节,以维持血清溶质的正常水平(Bourque,2008)。重量摩尔渗透压浓度定义为每公斤水溶质渗透摩尔数(mOsm / kg)(Dufour,1993),并且可以使用依赖于溶液的碰撞性质的不同技术来测量。实验室设置中最常用的是蒸气压和凝固点渗透压测量法,它们相对快速且易于执行。凝固点渗透压是优选的,因为它对溶解中可能存在的挥发性化合物(例如醇)不敏感。血清或血浆渗透压的测量在许多病症和疾病方面具有临床意义,包括高钠血症,糖尿病酮症酸中毒和不良抗生素综合征(Ellison,2013; Lupsa和Inzucchi,2013; Reddi,2013)。在本协议中,我们描述了使用冰点渗透压测定技术测量大鼠血清渗透压的方法,原先在我们以前在败血症中的渗透调节扰动研究中概述(Stare等,2015)。
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Isolation and Primary Cell Culture of Mouse Dorsal Root Ganglion Neurons
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Author:
Date:
2016-04-05
[Abstract] We here provide a detailed protocol for the isolation and culture of primary mouse sensory neurons. The cell bodies of sensory afferent pseudounipolar neurons are located in dorsal root ganglia (DRGs) along the vertebral column. Dissected mouse DRGs can be dissociated into single cells by enzymatic digestion to obtain primary cultures of mouse sensory neurons as performed in the studies reported by Khaminets et al. (2015).
[摘要] 我们在这里提供了详细的协议,用于隔离和培养的主要小鼠感觉神经元。 感觉传入假性极化神经元的细胞体位于沿着脊柱的背根神经节(DRG)中。 解离的小鼠DRG可以通过酶消化解离成单个细胞,以获得小鼠感觉神经元的原代培养物,如Khaminets等人(2015)报道的研究中所进行的。
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