BALB-c小鼠黑色素瘤B16移植肿瘤模型建立.doc

BALB/c小鼠黑色素瘤B16移植肿瘤模型建立 　【摘要】 ［目的］建立宜于进行早期观察及测量的小鼠黑色素瘤模型［方法］将不同数量的处于对数生长期的B16细胞株黑色素瘤细胞接种于BALB/c小鼠背部及足垫皮下，建立移植肿瘤动物模型.观察小鼠肿瘤的生长特点，应用病理学和免疫组织化学染色方法进行鉴定，归纳总结接种细胞数与肿瘤发生率的关系及肿瘤生长对小鼠生活状态的影响［结果］接种5d后BALB/c小鼠皮肤浅表部位相继出现黑色的点或线状改变，发展为结节状改变直至肿瘤破溃、出血、死亡或破溃、出血、结痂、愈合，部分小鼠出现肿瘤消退.在接种细胞数大于5×105个时BALB/c小鼠的肿瘤模型全部建立成功［结论］B16细胞株黑色素瘤细胞接种于BALB/c小鼠的肿瘤模型成功率与接种细胞数成正比，它具有建立肿瘤模型成功率高，观察及测量方便等优点，接种部位以背部皮下为宜. 【关键词】 黑色素瘤 模型 动物 小鼠 ABSTRACT:OBJECTIVETo establish neoplasm transplantation models of melanoma in BALB/c miceMETHODSB16 melanoma cells in logarithmic growth phase were inoculated into BALB/c mice’s back hypodermis and feet with different concentration to establish neoplasm transplantation of melanoma. The growth characteristics of transplanted melanoma in mice were observed and the tumor properties were identified by histopathology and immunohistochemistry. The relations between the number of B16 cells inoculated and tumorigenetic rate, and the influence of tumorigenesis for mice were observedRESULTSFive days later, black spots and linear changes were gradually seen in BALB/c mice’s back skin, feet or ankle, and then they developed to tubercles, ulceration, hemorrhage and death. The tumorigenetic rate in inoculating with 5×105 B16 cells was near 100%CONCLUSIONThe tumorigenetic rate has a positive relation to the number of inoculated B16 cells, and is high in BALB/c mice inoculated the B16 cells into back hypodermis. Key words:melanoma;models, animal;mice 恶性黑色素瘤是来源于黑色素细胞的恶性肿瘤，是起源于神经嵴的黑色素细胞恶性肿瘤［1］，近年来发病率呈逐年上升的趋势，目前尚无有效的治疗方法［2，3］.本实验通过给接种不同数量的黑色素瘤细胞，建立了便于早期观察的黑色素瘤动物模型，并筛选出最佳接种细胞数及接种部位，为进一步进行黑色素瘤的治疗实验研究奠定基础. 1 材料与方法 1.1 材料 实验动物取清洁级近交系BALB/c小鼠，购于中国军事医科大学实验动物中心，雄性，6，7周龄，体重为17～19g.仪器：二氧化碳细胞培养箱（MCO17AC）为日本SANYO公司产品.试剂：S100抗体及SP试剂盒分别为美国Santa Cruz公司及ZYMED公司产品. 1.2 方法 1.2.1 细胞培养 B16黑色素瘤细胞株购于上海细胞生物研究所，常规培养于RPMI1640 100mL/L胎牛血清加双抗（青霉素100U/mL，链霉素100mg/L）的培养液内，置于37℃，50mL/L二氧化碳培养箱中培养.稳定传代3，4代后，取对数生长期细胞，经2.5g/L胰酶消化后用无血清培养液重悬收集细胞，用无菌生理盐水调整细胞密度备用.用台盼蓝染细胞确定活细胞数在95%以上. 1.2.2 小鼠背部皮下肿瘤模型的建立 取BALB/c小鼠24只