目的生长动态的监测是肿瘤学重要的研究内容之一,现有的各种生长动态的监测方法存在诸多缺陷。文中研究活体成像技术(in vivo imaging technique,IVIT)在肿瘤生长动态监测中的应用。方法在18只Babl/C小鼠背部皮下注射1×106/mL B16-F10-luc-G5细胞100μL,建立种植瘤模型。肿块出现后每天测量肿瘤并计算肿瘤体积,并于第0、3、5、7、9、14天每次取3只小鼠以活体成像系统(in vivo imaging system,IVIS)检测肿瘤生物发光,检测后处死取肿瘤组织制成石蜡切片行病理学检查。结果所有小鼠接种成功肿瘤,第5天肿瘤可见。第5、7、9、14天肿瘤大小分别为[(2.86E+00)±1.21]、[(4.87E+00)±1.66]、[(9.27E+01)±6.31]、[(2.60E+02)±7.88]mm3;所有移植瘤生物发光均被IVIS检测到,第0、3、5、7、9、14天肿瘤平均实测光子数为[(6.35E+05)±7655]、[(1.12E+05)±1820]、[(1.62E+05)±2090]、[(2.40E+05)±3515]、[(1.18E+06)±11530]、[(2.23E+06)±17934]photon/(cm2·ser·s);移植瘤平均实测光子数与体积之间存在线性回归关系(R2=0.97);以IVIS监测结果绘制的肿瘤生长曲线,揭示了B16F10移植瘤生长动态特点。结论 IVIT可连续监测标记肿瘤的生长动态,准确灵敏、具有一定实用价值。
In vivo imaging system (IVIS) is a new and rapidly expanding technology, which has a wide range of applications in life science such as cell tracing. By counting the number of photons emitted from a specimen, IVIS can quantify biological events such as tumor growth. We used B16F10-luc-G5 tumor cells and 20 Babl/C mice injected subcutaneously with B16F10-luc-G5 tumor cells (1×106 in 100 μL) to develop a method to quantitatively analyze cells traced by IVIS in vitro and in vivo, respectively. The results showed a strong correlation between the number of tumor cells and the intensity of bioluminescence signal (R2=0.99) under different exposure conditions in in vitro assay. The results derived from the in vivo experiments showed that tumor luminescence was observed in all mice by IVIS at all days, and there was significant difference (P<0.01) between every two days from day 3 to day 14. Moreover, tumor dynamic morphology could be monitored by IVIS when it was in- visible. There was a strong correlation between tumor volume and bioluminescence signal (R2=0.97) by IVIS. In summary, we demonstrated a way to accurately carry out the quantitative analysis of cells using IVIS both in vitro and in vivo. The data indicate that IVIS can be used as an effective and quantitative method for cell tracing both in vitro and in vivo.
