van Dam P A, Watson J V, Lowe D G, Chard T, Shepherd J H
Department of Gynecologic Oncology, Saint Bartholomew's Hospital, London, England.
Cytometry. 1992;13(7):722-9. doi: 10.1002/cyto.990130708.
Five different protocols for the short-term preservation of cells used for multiparameter flow cytometric assay of tumour associated antigens (TAA) and DNA were assessed in cell suspensions prepared by mechanical disaggregation of 15 gynecological tumors. The protocols at 4 degrees C were 1) storage in buffer, 2) storage in 50% methanol, and 3) storage in buffer after formalin fixation. Tissues were also cryopreserved as cell suspensions and tissue blocks. When the TAA expression and DNA histograms of the preserved cells were compared with those in fresh cell suspensions, cryopreservation was found to be the best method: TAA expression was well preserved and there was a good correlation between TAA expression and the quality of the DNA histograms, respectively, in fresh and cryopreserved cells (RS: 0.82-0.91, P less than 0.001 for all TAAs). The cell suspensions preserved at 4 degrees C all showed a significant increase in background fluorescence (P less than 0.05) and a reduction in the TAA specific fluorescence (P less than 0.011). Methanol fixation was better than buffered formalin for the proteins studied, though both gave significantly worse results than cryopreservation. The quality of these cell suspensions and the correlation with TAA measurements in fresh cell suspensions deteriorated progressively with time, particularly if they were stored more than a week.
在通过机械解离15例妇科肿瘤制备的细胞悬液中，评估了五种用于短期保存用于肿瘤相关抗原（TAA）和DNA多参数流式细胞术检测的细胞的不同方案。4℃下的方案为：1）保存在缓冲液中；2）保存在50%甲醇中；3）福尔马林固定后保存在缓冲液中。组织也作为细胞悬液和组织块进行冷冻保存。当将保存细胞的TAA表达和DNA直方图与新鲜细胞悬液中的进行比较时，发现冷冻保存是最佳方法：TAA表达保存良好，新鲜细胞和冷冻保存细胞中TAA表达与DNA直方图质量之间分别具有良好的相关性（相关系数：0.82 - 0.91，所有TAA的P均小于0.001）。4℃保存的细胞悬液均显示背景荧光显著增加（P小于0.05）且TAA特异性荧光降低（P小于0.011）。对于所研究的蛋白质，甲醇固定优于缓冲福尔马林，不过二者的结果均明显差于冷冻保存。这些细胞悬液的质量以及与新鲜细胞悬液中TAA测量值的相关性随时间逐渐变差，尤其是如果保存超过一周。
