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A novel bioluminescent tumor model of human renal cancer cell lines: an in vitro and in vivo characterization.

机译:人肾癌细胞系的新型生物发光肿瘤模型:体外和体内表征。

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PURPOSE: Bioluminescent imaging permits sensitive in vivo detection and quantification of cells engineered to emit light. We developed a bioluminescent human renal cancer cell line for in vitro and in vivo studies. MATERIAL AND METHODS: The 2 human renal cell carcinoma cell lines SN12-C and SN12-L1 were stably transfected to constitutively express luciferase using a retroviral shuttle. The bioluminescent signal was correlated with tumor cell numbers in vitro. Parental and transfected cells were compared by growth kinetics and histology. Tumor burden after heterotopic injection in immune deficient mice was monitored up to 39 days. The kinetics of the bioluminescent signal was evaluated for 1 to 60 minutes following luciferin injection. RESULTS: Bioengineered renal cancer cell lines stably expressed luciferase. The growth kinetics of the cells in vitro and the histology of tumors resulting from implantation of these cells were unaffected by retroviral transfection with the luciferase gene. As few as 1,000 cells could be reliably detected. The intensity of the bioluminescent signal correlated with the number of tumor cells in vitro. Photon emission in vivo and ex vivo correlated significantly with tumor weight at sacrifice. After intraperitoneal injection of luciferin there was a time dependent change in the intensity of the bioluminescent signal with maximum photon emission at 20 minutes (optimal 17 to 25). CONCLUSIONS: Luciferase transfected human renal cancer lines allow reliable, rapid, noninvasive and longitudinal monitoring of tumor growth in vivo. The ability to assess tumor development in vivo with time is economical and effective compared to end point data experiments.
机译:目的:生物发光成像可以对体内工程发光的细胞进行灵敏的体内检测和定量。我们开发了一种生物发光的人类肾癌细胞系,用于体外和体内研究。材料与方法:使用逆转录病毒穿梭病毒将2种人肾癌细胞系SN12-C和SN12-L1稳定转染以组成型表达萤光素酶。生物发光信号与体外肿瘤细胞数量相关。通过生长动力学和组织学比较亲代和转染细胞。在免疫缺陷小鼠中异位注射后的肿瘤负荷监测长达39天。荧光素注射后,在1至60分钟内评估生物发光信号的动力学。结果:生物工程肾癌细胞系稳定表达荧光素酶。荧光素酶基因逆转录病毒转染不会影响细胞的体外生长动力学和因植入这些细胞而导致的肿瘤组织学。可以可靠地检测到多达1,000个细胞。生物发光信号的强度与体外肿瘤细胞的数量有关。体内和离体的光子发射与处死时的肿瘤重量显着相关。腹膜内注射荧光素后,生物发光信号强度随时间变化,在20分钟时光子发射最大(最佳17到25)。结论:荧光素酶转染的人肾癌系允许可靠,快速,无创和纵向监测体内肿瘤的生长。与终点数据实验相比,评估随时间变化的体内肿瘤发展的能力既经济又有效。

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