Maley D T, Simon P
Medical Products Department, E.I. DuPont de Nemours, Inc., Glenolden, PA 19036.
J Immunol Methods. 1990 Nov 6;134(1):61-70. doi: 10.1016/0022-1759(90)90112-9.
We describe a method for standardizing cytotoxicity assays by the use of cryopreserved, fluorescently labeled target cells. The cells are labeled in batches with the fluorogenic element europium (Eu) and frozen in multiple aliquots. Replicate aliquots can be thawed on different days and used for cytotoxicity assays. Thawed pre-labeled cells from the same batch are killed reproducibly, allowing cytotoxicity assays to be standardized. The target cells need only be thawed, washed, counted for viability and used. The availability of pre-labeled, cryopreserved targets facilitates the assessment of cytotoxic activity of individual donors at different times. The method also simplifies the use of large panels of target cell types. By using this procedure it is not necessary to maintain active cultures of target cells, nor is it necessary to use and dispose of radioisotopes. These pre-labeled target cells can be used in various assays where the result is a destroyed target cell such as cell-mediated cytotoxicity, complement mediated cytotoxicity, or antibody-dependent cellular cytotoxicity.
我们描述了一种通过使用冷冻保存的、荧光标记的靶细胞来标准化细胞毒性测定的方法。细胞分批用荧光元素铕（Eu）标记，并冷冻成多个等分试样。重复的等分试样可在不同日期解冻并用于细胞毒性测定。来自同一批次的解冻预标记细胞可被可重复地杀死，从而使细胞毒性测定得以标准化。靶细胞只需解冻、洗涤、计数活力并使用即可。预标记的、冷冻保存的靶细胞的可用性有助于在不同时间评估个体供体的细胞毒性活性。该方法还简化了大量靶细胞类型的使用。通过使用此程序，无需维持靶细胞的活跃培养物，也无需使用和处置放射性同位素。这些预标记的靶细胞可用于各种测定，其中结果是靶细胞被破坏，如细胞介导的细胞毒性、补体介导的细胞毒性或抗体依赖性细胞毒性。
