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目的:A20细胞是来源于同系Balb/c小鼠大B细胞淋巴瘤的细胞系,能通过尾静脉接种建立小鼠B细胞播散型淋巴瘤的模型,但由于该细胞系缺乏细胞表面特异性标志而难于监测。本研究使用增强型绿色荧光蛋白(EGFP)标记A20细胞,试图建立易于监测的小鼠B细胞播散型淋巴瘤模型。方法:用含增强型绿色荧光蛋白(EGFP)的慢病毒载体将标记基因EGFP转入A20细胞,通过流式分选出EGFP+的A20细胞,体外培养后通过尾静脉注射接种于同系Balb/c小鼠体内,用流式细胞仪监测其外周血EGFP+细胞的百分率。当小鼠出现消瘦、毛发竖立、嗜睡等体征时,将小鼠处以安乐死;取动物脏器行石蜡包埋、病理切片、HE染色。结果:尾静脉注射1×106细胞于6只Balb/c小鼠体内,接种后15天可在外周血中检测到EGFP+细胞,平均生存时间为29.6±0.8天;在肝脏、脾脏、脊椎和淋巴结等多脏器成瘤,流式检测瘤细胞EGFP表达阳性。结论:经尾静脉注射接种A20细胞可建立小鼠B细胞播散型淋巴瘤模型,A20细胞经含EGFP的慢病毒标记后易于通过流式进行监测,为通过动物体内试验评价靶向治疗的疗效提供了保证。
OBJECTIVE: A20 cells are cell lines derived from large B-cell lymphoma of syngeneic Balb / c mice and can be used to establish a model of mouse B-cell disseminated lymphoma through tail vein inoculation. However, due to the lack of cell surface specificity Logo and difficult to monitor. In this study, A20 cells were labeled with enhanced green fluorescent protein (EGFP) in order to establish a mouse B cell disseminated lymphoma model that is easy to monitor. Methods: EGFP was transfected into A20 cells by lentiviral vector containing enhanced green fluorescent protein (EGFP). EGFP + A20 cells were separated by flow cytometry and cultured in vitro. In vivo, the percentage of EGFP + cells in peripheral blood was monitored by flow cytometry. When mice showed weight loss, hair erection, lethargy and other signs, the mice were euthanized; take animal paraffin-embedded, pathological sections, HE staining. Results: 1 × 106 cells were injected into the tail vein of 6 Balb / c mice via tail vein. EGFP + cells were detected in peripheral blood 15 days after inoculation, with an average survival time of 29.6 ± 0.8 days. In liver, spleen, spine and lymph nodes Such as multiple organ tumor, flow cytometry positive EGFP expression. CONCLUSION: A mouse B-cell disseminated lymphoma model can be established by inoculation of A20 cells via tail vein injection. A20 cells can be easily monitored by flow cytometry after being labeled with EGFP-containing lentivirus. In order to evaluate the efficacy of targeted therapy in animal experiments Provided a guarantee.