| Primary Explosive Blast-induced Traumatic Brain Injury Model in PC12 Cell Culture
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Author:
Date:
2016-08-20
[Abstract] While it is understood that structural damage occurs at the cellular level from the traumatic brain injury event, the effect on functional activity remains largely unknown. Simplified models such as in vitro models of primary explosive blast are critically needed to deconvolute mechanisms of cellular damage. This protocol details an in vitro indoor experimental system setup (Zander et al., 2015) using real military explosive charges to more accurately represent battlefield blast exposure, and probe the effects of primary explosive blast on dissociated neurons.
[摘要] 细胞毒性CD8 + T细胞能够通过其T细胞受体（TCR）与主要组织相容性复合物（MHC）分子呈递的小免疫原性肽（抗原）之间的特异性相互作用特异性识别和杀死靶细胞。抗原特异性细胞毒性T细胞的抗原识别能力和体外裂解活性可以在所谓的铬51（<51> Cr）释放测定中功能性评估，其是几乎50年前在我们的机构中发展起来的（Brunner等人，1968年）。放射性标记的内源性抗原呈递缺陷的细胞[例如，用于抗原呈递（TAP）缺陷型T2细胞的转运蛋白]，并用感兴趣的MHC稳定转染（例如 ，HLA-A2 sup + +）通常在这个4小时测定期间用作靶标。或者，内源性呈递免疫原性抗原的Cr标记的病毒感染或肿瘤细胞系可以作为靶细胞（例如，用于评估肿瘤识别）。 在肽滴定测定（部分A）中，用抗原性肽的系列稀释物对放射性标记的靶细胞进行脉冲，并在效应物（例如，CD8 + T细胞克隆）与靶细胞（ Cr-T2细胞）比（E:T）为10:1的混合物在96孔V型底板中37℃温育4小时。在肿瘤杀伤试验（B部分）中，将细胞毒性CD8 ...
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| Neuron Culture from Mouse Superior Cervical Ganglion
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Author:
Date:
2014-01-20
[Abstract] The rodent superior cervical ganglion (SCG) is a useful and readily accessible source of neurons for studying the mechanisms of sympathetic nervous system (SNS) development and growth in vitro. The sympathetic nervous system (SNS) of early postnatal animals undergoes a great deal of remodeling and development; thus, neurons taken from mice at this age are primed to re-grow and establish synaptic connections after in situ removal. The stereotypic location and size of the SCG make it ideal for rapid isolation and dissociation. The protocol described here details the requirements for the dissection, culture and differentiation of SCG neurons. The protocol is suitable for culturing neurons from late embryonic gestation to approximately postnatal day 3. The culture technique ...
[摘要] 啮齿动物上级子宫颈神经节（SCG）是一种有用且易于获取的神经元来源，用于研究交感神经系统（SNS）体外发育和生长的机制。 早期出生后动物的交感神经系统（SNS）经历了大量的重塑和发育; 因此，在这个年龄的小鼠中取出的神经元被引发再生长并在原位移除后建立突触连接。 SCG的刻板位置和尺寸使其成为快速隔离和解离的理想选择。 这里描述的方案详述了SCG神经元的解剖，培养和分化的要求。 该方案适用于从晚期胚胎妊娠至大约出生后第3天培养神经元。下面讨论的培养技术利用玻璃盖玻片进行固定细胞的显微镜检查。
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