■细胞处理设施在处理过程中面临环境细菌污染生物安全柜的风险,人工处理自体细胞产品会导致污染。我们提出了一种基于风险和证据的交叉污染清洁方法,强调蛋白质和DNA。
■通过使用荧光强度测量湿和干燥介质来评估培养基的转变和残留风险。在紫外线(UV)照射后,分析了含有HT-1080细胞的干燥培养基中的残留蛋白质和DNA,擦拭,和消毒剂处理。
■潮湿条件显示出更高的向蒸馏水(DW)的过渡,而干燥条件导致SUS304板上较高的残留量。检查了残留培养基的各种清洁方法,包括苯扎氯铵和缓蚀剂(BKC+I)和DW擦拭,与其他方法相比,这表明残留蛋白质和DNA显着降低。此外,这些清洁方法测试了含有细胞的残留培养基,用BKC+I和DW擦拭导致无法检测到的细胞数量。然而,在某些情况下,蛋白质和DNA仍然存在。
■该研究比较了细胞产品中蛋白质和DNA的清洁方法,揭示它们的优点和缺点。过氧乙酸(PAA)被证明对核酸有效,但对蛋白质无效,而紫外线照射对蛋白质和DNA都无效。擦拭成为最有效的方法,尽管可追溯性仍然具有挑战性。然而,用ETH擦拭无效,因为它引起蛋白质固定。了解这些清洁方法的特点对于制定有效的污染控制策略至关重要。
UNASSIGNED: Cell-processing facilities face the risk of environmental bacteria contaminating biosafety cabinets during processing, and manual handling of autologous cell products can result in contamination. We propose a risk- and evidence-based cleaning method for cross-contamination, emphasizing proteins and DNA.
UNASSIGNED: The transition and residual risks of the culture medium were assessed by measuring both wet and dried media using fluorescence intensity. Residual proteins and DNA in dried culture medium containing HT-1080 cells were analyzed following ultraviolet (UV) irradiation, wiping, and disinfectant treatment.
UNASSIGNED: Wet conditions showed a higher transition to distilled water (DW), whereas dry conditions led to higher residual amounts on SUS304 plates. Various cleaning methods for residual culture medium were examined, including benzalkonium chloride with a corrosion inhibitor (BKC + I) and DW wiping, which demonstrated significantly lower residual protein and DNA compared to other methods. Furthermore, these cleaning methods were tested for residual medium containing cells, with BKC + I and DW wiping resulting in an undetectable number of cells. However, in some instances, proteins and DNA remained.
UNASSIGNED: The study compared cleaning methods for proteins and DNA in cell products, revealing their advantages and disadvantages. Peracetic acid (PAA) proved effective for nucleic acids but not proteins, while UV irradiation was ineffective against both proteins and DNA. Wiping emerged as the most effective method, even though traceability remained challenging. However, wiping with ETH was not effective as it caused protein immobilization. Understanding the characteristics of these cleaning methods is crucial for developing effective contamination control strategies.