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目的探讨冻存H22肝癌细胞应用于卡介苗抑瘤小鼠模型的可行性。方法制备冻存H22细胞,分别将冻存10和83 d的H22细胞复苏,计算细胞存活率;将高(7.5×106个/ml)、中(5.0×106个/ml)、低(2.5×106个/ml)剂量的新复苏的H22细胞与1 mg/ml治疗用卡介苗混合后经左侧背部皮下免疫BALB/c小鼠,0.1 ml/只,对照组只接种相应浓度的H22细胞,接种后30 d,称量小鼠皮下瘤体重量,并计算试验组的抑瘤率,试验重复2次。结果冻存10和83 d的H22细胞复苏后的存活率分别为85.45%和82.25%。所有对照组小鼠接种H22细胞后均有肿瘤形成,成瘤率均为100%;同一次试验中,各试验组瘤体重量均明显小于同剂量对照组(P<0.05);同一次试验中,各试验组间瘤体重量差异无统计学意义(P>0.05);不同次试验中,同剂量对照组间瘤体重量差异均有统计学意义(P<0.05);各试验组的抑瘤率均大于60%。结论冻存H22细胞可满足抑瘤试验需要,但试验稳定性仍需改进。
Objective To investigate the feasibility of using cryopreserved H22 hepatocarcinoma cells in BCG-resistant mice. Methods H22 cells were cryopreserved, resuscitation of H22 cells at 10 and 83 days after cryopreservation were performed, and cell viability was calculated. High (7.5 × 106 cells / ml) medium (5.0 × 106 cells / ml) 106 BALB / c mice were immunized subcutaneously with 0.1 mg / ml of H22 cells with a dose of 106 / ml. The control group was inoculated only with the corresponding concentration of H22 cells and inoculated After 30 days, the weight of subcutaneous tumor was weighed and the inhibition rate of the test group was calculated. The experiment was repeated twice. Results The survival rates of H22 cells cryopreserved for 10 and 83 days after resuscitation were 85.45% and 82.25%, respectively. All the control mice developed tumor formation after inoculation of H22 cells, with tumor formation rates of 100%. In the same experiment, the tumor weight of each experimental group was significantly smaller than that of the same dose control group (P <0.05); in the same experiment (P> 0.05). In different times, the difference of tumor weight between the same dosage of control group was statistically significant (P <0.05) Rate of more than 60%. Conclusion Cryopreservation of H22 cells can meet the need of antitumor test, but the test stability still needs improvement.