Roth Susanne Pauline, Liso Giulia, Brehm Walter, Wagner Bettina, Schnabel Christiane Liliane, Troillet Antonia
Veterinary Teaching Hospital, Department for Horses, Faculty of Veterinary Medicine, Leipzig University, Leipzig, Germany.
Saxonian Incubator for Clinical Translation (SIKT), Leipzig University, Leipzig, Germany.
Front Vet Sci. 2025 May 27;12:1588240. doi: 10.3389/fvets.2025.1588240. eCollection 2025.
The anti-inflammatory and regenerative effects of autologous conditioned serum (ACS) in joint-associated disorders are presumably mediated by the biomolecules it contains, such as cytokines and chemokines. ACS is commonly used in equine practice after storage. Data regarding the influence of storage conditions of ACS on cytokine and chemokine concentrations are not available.
To evaluate the effect of commonly used storage conditions on selected cytokine and chemokine concentrations in equine ACS.
From 10 horse patients with clinically indicated ACS preparation six ACS aliquots each were stored under different storage conditions. The samples were evaluated after storage at room temperature (rt) or at refrigerator temperature (6-8°C) for 6 h. Another set of samples was stored frozen at -20°C for 7 months. After this storage under clinically relevant conditions all samples were frozen at -80°C, and compared to an aliquot immediately frozen at -80°C. Mediator concentrations were determined by fluorescent bead-based multiplex assays using monoclonal antibodies that are specific for the respective analyzed molecule: interferon (IFN)-γ, interleukin (IL)-17A, IL-4, IL-10, and IFN-α (equine cytokine 5-plex assay) as well as CC-chemokine ligand 2 (CCL2), CCL3, CCL5, CCL11, tumor necrosis factor (TNF)-α, and IL-1β (equine chemokine 6-plex assay) were analyzed.
Concentrations of IFN-γ and IL-10, as well as the chemokines analyzed, showed marked interindividual differences. The cytokines IL-17A, IL-4, and IFN-α were not detected in any of the samples analyzed. Concentrations of selected cytokines, and chemokines were similar between the defined storage conditions.
Concentrations of selected cytokines, chemokines, and IL-1ra in equine ACS were not influenced by the defined storage conditions studied (rt, refrigerated, frozen).
自体条件血清（ACS）在关节相关疾病中的抗炎和再生作用可能是由其所含的生物分子介导的，如细胞因子和趋化因子。ACS在储存后常用于马的临床实践。关于ACS储存条件对细胞因子和趋化因子浓度影响的数据尚无可用信息。
评估常用储存条件对马ACS中选定细胞因子和趋化因子浓度的影响。
从10例有临床指征需制备ACS的马患者中，每份ACS等分试样分别储存在不同的储存条件下。样品在室温（rt）或冰箱温度（6 - 8°C）下储存6小时后进行评估。另一组样品在-20°C下冷冻储存7个月。在这些临床相关条件下储存后，所有样品均在-80°C下冷冻，并与立即在-80°C下冷冻的等分试样进行比较。使用对各自分析分子具有特异性的单克隆抗体，通过基于荧光微球的多重分析来测定介质浓度：干扰素（IFN）-γ、白细胞介素（IL）-17A、IL-4、IL-10和IFN-α（马细胞因子5重分析）以及CC趋化因子配体2（CCL2）、CCL3、CCL5、CCL11、肿瘤坏死因子（TNF）-α和IL-1β（马趋化因子6重分析）。
IFN-γ和IL-10的浓度以及所分析的趋化因子显示出明显的个体差异。在所分析的任何样品中均未检测到细胞因子IL-17A、IL-4和IFN-α。选定的细胞因子和趋化因子在规定的储存条件下浓度相似。
马ACS中选定的细胞因子、趋化因子和IL-1ra的浓度不受所研究的规定储存条件（室温、冷藏、冷冻）的影响。
