B16-F10-luc-G5黑色素瘤细胞生物学特性的研究

目的：探讨B16-F10-luc-G5黑色素瘤细胞的生物学特性.方法：倒置荧光显微镜下连续观察B16-F10-luc-G5细胞生长动态.流式细胞术检测冻存复苏后、培养基漂浮细胞、胰酶消化损伤后的细胞死亡率以评价其对实验损伤的耐受性.1×104/孔B16-F10-luc-G5细胞按1∶2梯度稀释至0.78×102/孔,加入底物荧光素后检测其生物发光特性.Babl/C和C57 bl/6雄性小鼠各10只接种该细胞,目测肿瘤生长速度和活体成像监测Babl/C小鼠移植瘤生长动态以评价其成瘤特性.结果：B16-Fl0-luc-G5细胞生长动态符合典型B16-F10细胞特性,无自发及激发荧光.B16-F10-luc-G5细胞冻存、漂浮细胞及对数生长期细胞消化后死亡率分别为23.8％、35.8％和4.8％.B16-F10-luc-G5细胞数与实测平均光子数之间存在线性回归关系,检测光子数可反映细胞数目.两组小鼠移植成瘤率均为100％,平均肿瘤出现时间差异显著（t =9.05,P＜0.05）,移植瘤病理符合B16黑色素瘤典型生长特点;活体成像监测可灵敏监测移植瘤生长动态.结论：B16-F10-luc-G5细胞具有生长快、对各种实验操作耐受性好、标记生物发光基因易追踪等优点,且其一般特性与B16-F10细胞基本相同,是肿瘤学研究良好的实验材料.

Cytobiological characters of mouse melanoma cell lines B16-F10-luc-G5

Objective：To study the biological characteristics of B16 - FIO mouse melanoma cell labeling luc gene （ B16 - F10 - luc - GS）. Methods：Tumor dynamic morphology of B16 - F10 - luc - G5 was observed under fluores- cence microscope. Activity after various kinds of experimental treatment was detected by flow cytometry. B16 - FIO - luc - G5 cells were diluted from 10,0OO cells to 78 cells and then the cell plate was imaged using IVIS system to eval- uate its 1 characteristics. Ten Babl/C and C57bl/6 mice with subcutaneously injected tumor cells （ 1 ~ 105 in lOOp.l）, then the growth curves and pathologic changes of transplanted tumor were compared. Luminescences of transplanted tumor were sequentially detected by IVIS. Results：Tumor dynamic morphology of B16 - FIO - luc - G5 was nearly the same as typical B16 -F10 cell line. Apoptosis of B 16 - F10 - luc - G5 cells under several condi- tions weredetected by flow cytometry. A strong correlation between number of tumor cells and bioluminescence （r = 0. 998）was also demonstrated by IVIS. There was significant difference （P 〈 0.01 ）of tumor growth between two groups. Tumor dynamic morphology could still be detected by IVIS even when its cell was invisible. Conclusion ： B16 -F10 -luc -G5 cell labeled with luc gene, had typical biological characteristics of melanoma cell, and may be rec- ommended as a useful material for in vivo research because it could be traced by imaging technique.