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目的建立RAG2/IL2RG双基因缺陷的CRG小鼠杂交群体。方法将RAG2基因缺陷小鼠与IL2RG基因缺陷小鼠分别进行繁殖,选取RAG2基因缺陷雄鼠RAG2(-/-)与IL2RG基因缺陷雌鼠IL2RG(-/-)进行配对,培育杂交后代,通过PCR扩增基因组RAG2和IL2RG基因进行鉴定;应用流式细胞仪分析检测RAG2/IL2RG双基因缺陷小鼠外周血T细胞(CD3+)、B细胞(CD19+)和NK细胞(CD49b+)含量;并测定8—12周龄RAG2/IL2RG双基因缺陷小鼠血液生理生化及主要脏器重量指标。结果成功繁育RAG2基因缺陷小鼠和IL2RG基因缺陷小鼠,筛选获得稳定的RAG2/IL2RG双基因缺陷小鼠并成功保种和扩群;该双基因缺陷小鼠外周血淋巴细胞中T细胞、B细胞,NK细胞比例显著降低(P<0.05);与相同周龄野生型C57BL/6相比9项血清生化指标无明显变化(P>0.05);18项血液生理指标中红细胞(RBC)和血红蛋白(HGB)含量显著降低(P<0.05);白细胞(WBC)、淋巴细胞数(LYMPH)、淋巴细胞比率(LYM)、单核细胞、中性细胞数(NEUT)降低极显著(P<0.01);脾脏重量显著均低于野生型C57BL/6小鼠(P<0.05);人肝肿瘤传代细胞移植于RAG2/IL2RG双基因缺陷小鼠后,肿瘤移植成功率和生长速率均明显高于亲本单基因缺陷小鼠(P<0.05),成功获得了临床肝癌病人肿瘤组织的异种移植模型。结论成功构建筛选出RAG2/IL2RG双基因缺陷小鼠杂交群体。
Objective To establish a hybrid population of RAG2 / IL2RG double gene defect CRG mice. Methods RAG2 gene-deficient mice and IL2RG gene-deficient mice were propagated separately. The RAG2 gene-deficient male RAG2 (- / -) and IL2RG gene-deficient female IL2RG (- / - The genomic RAG2 and IL2RG genes were amplified and identified. The levels of CD3 +, CD19 + and CD49b + were detected by flow cytometry in peripheral blood of RAG2 / IL2RG double gene-deficient mice. Blood Physiological and Biochemical Parameters of 12-week-old RAG2 / IL2RG Double Gene-deficient Mice and Indicators of the Weight of Major Organs. Results The RAG2 gene-deficient mice and IL2RG gene-deficient mice were successfully bred and the stable RAG2 / IL2RG double gene-deficient mice were screened and successfully maintained for seedling expansion and expansion. The T cells in B-deficient mouse peripheral blood lymphocytes, B (P <0.05). There were no significant changes in the serum biochemical parameters of 9 serums in the same weeks of age compared with the wild type C57BL / 6 (P> 0.05); 18 blood physiological indexes of red blood cells (RBC) and hemoglobin (P <0.05). The WBC, LYMPH, LYM, monocytes and NEUT decreased significantly (P <0.01) ; The weight of spleen was significantly lower than that of wild-type C57BL / 6 mice (P <0.05). The success rate and growth rate of tumor transplants were significantly higher in human hepatoma cell line transplanted in RAG2 / IL2RG double gene-deficient mice Gene-deficient mice (P <0.05) successfully obtained the xenograft model of tumor tissue in clinical liver cancer patients. Conclusion The hybrids of RAG2 / IL2RG double gene-deficient mice were successfully constructed.