目的探讨Hepa1-6接种于C57BL/6j小鼠皮下建立适合免疫学研究的肿瘤模型的可行性。方法Hepa1-6细胞培养传代3~4次后,收集并浓缩为3个浓度梯度:5×10^6/ml,1×10^7/ml和2×10^7/ml。按0.1ml/鼠的量接种于C57BL/6j小鼠皮下。调查每组的皮下肿瘤形成率。接种7d后。每3天测量肿瘤的最大直径及横径。计算肿瘤体积、生长速率及倍增时间。再得出生长曲线。观察荷瘤鼠与未接种鼠的生存时间;HE染色观察肿瘤病理学特点。结果Hepa1-6细胞培养24~36h。即可进行1:3的细胞传代。A,B组以及C组的成瘤率分别为10%(1/10)。20%(2/10)和90%(9/10)。最早扪及米粒大小的肿瘤结节的时间分别为19,10和7d。荷瘤小鼠生存时间为49~73d,中位生存时间为57.5d。平均生存时间为(61.4±12.2)d;非荷瘤小鼠生存时间为74~103d。中位生存时间为85d,平均生存时间为(85.6±15.1)d(t=3.45,P〈0.05)。肿瘤HE染色见肿瘤细胞排列不规则,多见病理性核分裂。结论以C57BL/6j为实验鼠,按2×10^6/小鼠的细胞量,皮下接种Hepa1-6细胞。能成功建立适合进行微波消融相关的免疫学层面研究的皮下肝癌模型。
Objective To establish a hepatoma model suitable for immunologic research by injection of hepa1-6 cell into C57BL/6j mice. Methods Hepa1-6 cells were collected and condensed as 5 × 10^6/ml, 1 × 10^7/ml and 2 × 10^7/ml, before the cells were injected subcutaneously with a dose of 5 × 10^5/mouse(group A, 10 mice), 1 × 10^6/mouse (group B, 10 mice) and 2 × 10^6/mouse(group C, 10 mice). The tumor was monitored for its generation rate, diameter and gross tumor volume. By using the above data, the growth rate and doubing time of tumor were calculated, and growth curve was generated. The survival time was observed in mice bearing tumor and those without tumor. Tumors were removed and stained with HE. Its pathologic characteristics were observed. Results Hepa1-6 cell could passage as 1:3 after 24 -36 hours. The tumor generation rate was 10% (1/10) in group A, 20% (2/10) in group B and 90% (9/10) in group C. The times when the tumor mass were palpable were 7 days, 10 days and 19 days. There was significant difference in average survival time (61.4 ± 12.2 d vs 85.6 ± 15. 1 d, t = 3.45, P 〈 0. 05 ). between tumor - bearing and healthy mice. Under microscope, the tumor structure was unregular and there were a lot of pathological nuclear division. Conclusion Subcutaneous hepatoma model suitable for thermo - ablation can be successfully established by subcutaneous inoculation of Hepal - 6 cell on C57BL/6j mice.