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目的探讨溴脱氧尿嘧啶核苷（ＢＵｄＲ）标记ＢＵｄＲ／ＤＮＡ双参数流式细胞仪（ＦＣＭ）测定肿瘤潜在倍增时间Ｔｐｏｔ的方法，比较ＢＵｄＲ在体（ｉｎｖｉｖｏ）、离体（ｉｎｖｉｔｒｏ）标记结果的差异。材料与方法实验采用人鼻咽癌裸小鼠移植瘤模型分别进行ＢＵｄＲ在体、离体标记；离体标记时分别将样本制成组织块和细胞悬液在０．５ｍＭ、０．１ｍＭＢＵｄＲ的含２０％胎牛血清ＲＰＭＩ１６４０培养基、５％ＣＯ２、３７℃、ＣＯ２孵箱中开放培养不同时间后收集样本。在体标记：腹腔注射ＢＵｄＲ０．１ｍｇ／ｇ体重，６小时后处死小鼠取瘤剪碎，在７０％的冷乙醇固定冰箱保存；全部样本用ＦＩＴＣＰＩ染色后行ＦＣＭ测定。结果（１）本实验室的ＢＵｄＲ离体培养条件下，组织块标记较理想。（２）用ＢＵｄＲ在体标记法测定不同大小的肿瘤（直径０．３～１．２ｃｍ），结果表明：直径＜０．５ｃｍ的肿瘤的标记效果比较理想。（３）相同体积的肿瘤在体（ｎ＝９）、离体（ｎ＝７）标记两种不同标记方法的结果显示，相同体积的肿瘤用ＢＵｄＲ离体标记法时ＬＩ比在体标记低５３％，其它各项细胞动力学参数两者均有明显差异，但离体标记法的组内变异很小。结论本实验室成功建立了在体ＢＵｄＲ标记ＢＵ?
OBJECTIVE: To investigate the method of determining tumor potential doubling time Tpot by bromodeoxyuridine (BUdR)-labeled BUdR/DNA double-parameter flow cytometry (FCM), and to compare the difference in in vitro and in vivo results of BUdR. . Materials and Methods Experiments were carried out using the human nudeo-carcinoma nude mouse xenograft tumor model for in vivo and ex vivo labeling of BUdR. When ex vivo labeling, the samples were made into tissue blocks and cell suspensions containing 0.5mM, 0.1mM BUdR. 20% fetal bovine serum in RPMI1640 medium, 5% CO2, 37°C, and CO2 incubators were collected at different times after open culture. In vivo labeling: intraperitoneal injection of BUdR 0.1mg/g body weight, 6 hours after the mice were sacrificed and the tumors were smashed and stored in a 70% cold ethanol fixed refrigerator; all samples were stained with FITC-PI and then subjected to FCM. Results (1) The tissue block marker was ideal under the in vitro culture condition of BUdR in our laboratory. (2) Tumors of different sizes (0.3 to 1.2 cm in diameter) were measured by BudR in vivo. The results showed that tumors with a diameter of <0.5 cm had an ideal labeling effect. (3) The results of two different labeling methods for the same volume of tumors in vivo (n=9) and ex vivo (n=7) showed that the same volume of tumor was markedly lower by in vitro labeling with BUd than LI in body markers. %, there are significant differences in other cell kinetic parameters, but the intra-group variation of the in vitro labeling method is very small. Conclusion The laboratory has successfully established BUdR BU in vivo.