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目的 探索建立人肝癌PBL SCID嵌合模型的方法。方法 C57/BL SCID小鼠腹腔注射健康志愿者外周血淋巴细胞(huPBL) 2×107 (1 0 ml),同时右腋部皮下注射 5×105 (0 2 ml)HepG2细胞。第2、4周鼠尾取血ELISA法测定人 IgG含量,用流式细胞仪分析小鼠外周血单个核细胞中人淋巴细胞比例。4周MTT法测定小鼠脾细胞对 HepG2 细胞的杀伤能力,免疫组化检测小鼠脾脏中人淋巴细胞的存在。结果 模型成瘤率为100%,成瘤潜伏期12~18 d。2、4周时小鼠血中人IgG 分别达69 8μg/ml、125 9μg/ml,人淋巴细胞占单核细胞的比例分别为 10 5%和 3 5%。4周时免疫组化检测出人淋巴细胞分布于小鼠脾脏中,小鼠脾细胞对 HepG2 细胞的杀伤力为 20 3%。处死动物,常规病理切片见肿瘤生长。结论 SCID小鼠腹腔注射人淋巴细胞、皮下注射肝癌细胞可建立人肝癌PBL SCID嵌合模型。
Objective To explore a method to establish chimeric model of human hepatocellular carcinoma PBL SCID. Methods C57 / BL SCID mice were injected intraperitoneally with 2 × 107 (10 ml) of peripheral blood lymphocytes (huPBL) in healthy volunteers and 5 × 105 (0 2 ml) HepG2 cells in the right axilla. At the end of the second and fourth week, the amount of human IgG in the tail of mice was measured by ELISA, and the proportion of human lymphocytes in peripheral blood mononuclear cells was analyzed by flow cytometry. The killing ability of mouse spleen cells on HepG2 cells was determined by MTT assay at 4 weeks, and the presence of human lymphocytes in spleen of mice was detected by immunohistochemistry. Results The rate of tumor formation was 100% and the tumorigenic potential was 12-18 days. At the 2nd and 4th week, the human IgG in the blood of the mice reached 69 8 μg / ml and 125 9 μg / ml, respectively. The ratios of human lymphocytes to mononuclear cells were 105% and 35% respectively. At 4 weeks, the distribution of human lymphocytes in mouse spleen was detected by immunohistochemistry. The lethality of mouse spleen cells to HepG2 cells was 20 3%. Animals were sacrificed, routine pathology see tumor growth. Conclusion SCID mice were injected intraperitoneally with human lymphocytes and subcutaneously injected with hepatoma cells to establish the human hepatocellular carcinoma PBL SCID chimerism model.