Cytokine networks in cerebrospinal fluid (CSF) are important to our understanding of several neuroinflammatory diseases. Knowledge about optimal handling of samples is limited but important to minimize bias and reduce costs in CSF biomarker studies. The aim of this study was to examine the effect of storage temperature and time delay from CSF sample collection until freezing on the concentration of 11 different cytokines thought to be associated with chronic pain. CSF samples from 21 individuals undergoing hip or knee arthroplasty under spinal anesthesia were divided between two tubes. One tube was stored and centrifuged (within 30 min) at room temperature, and one tube was stored in ice water and centrifuged (within 30 min) at 4 °C. Each tube was split into six vials that were frozen at −80 °C, 0.5, 1, 2, 3, 4, or 5 h after collection. Cytokines were analyzed using a multiplex panel. A random effect panel data regression was conducted for each biomarker including the variables of storage temperature until freezing and time delay. Four cytokines had detectable levels: Fractalkine, monocyte chemoattractant protein 1(MCP-1), interleukine 6 (IL-6), and interleukine 8 (IL-8). There was no significant effect of storage temperature and time delay on MCP-1, IL-6, or IL-8 concentrations. Fractalkine concentration showed no clear trend. No concentration differences were observed between samples kept in ice water and those at room temperature except at the 3-h time point, and there was no overall significant effect of time delay on fractalkine concentration. We found no clear effect of storage temperature and time delay up to five hours from sample collection until freezing on the CSF concentrations of fractalkine, MCP-1, IL-6, or IL-8.

脑脊液 (CSF) 中的细胞因子网络对我们了解几种神经炎症疾病很重要。关于样本最佳处理的知识有限，但对于减少 CSF 生物标志物研究中的偏差和降低成本很重要。本研究的目的是检查储存温度和从 CSF 样本收集到冷冻的时间延迟对被认为与慢性疼痛相关的 11 种不同细胞因子浓度的影响。来自 21 名在脊椎麻醉下进行髋关节或膝关节置换术的个体的脑脊液样本被分成两个管子。一管在室温下储存并离心（30分钟内），一管在冰水中储存并在4°C下离心（30分钟内）。每管分成六个小瓶，在收集后 0.5、1、2、3、4 或 5 小时，在 -80 °C 下冷冻。使用多重面板分析细胞因子。对每个生物标志物进行随机效应面板数据回归，包括冷冻前的储存温度和时间延迟变量。四种细胞因子具有可检测水平：Fractalkine、单核细胞趋化蛋白 1 (MCP-1)、白介素 6 (IL-6) 和白介素 8 (IL-8)。储存温度和时间延迟对 MCP-1、IL-6 或 IL-8 浓度没有显着影响。Fractalkine 浓度没有显示出明显的趋势。除了在 3 小时的时间点外，在冰水中保存的样品和室温下的样品之间没有观察到浓度差异，并且时间延迟对 fractalkine 浓度没有整体显着影响。