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目的:评价表达人肝再生增强因子基因的HepG2细胞系的细胞培养上清及细胞裂解物的小鼠急性毒性和近期致瘤性。方法:SPF级昆明种小鼠18只,随机分为空白对照组、细胞培养上清组、细胞裂解物组,每组小鼠各6只,腹腔分别接种空白培养液、细胞培养上清、细胞裂解物0.5ml。连续14天,每天观察记录动物毒性反应,14d后宰杀小鼠,取血测血生化指标,及观察病理改变。结果:各组小鼠均存活。除对照组1例小鼠,细胞培养上清组1例小鼠,细胞裂解物组2例小鼠次日活动稍减少外,均未见异常反应。血液生化检测ALT、AST、AFP、TBIL无明显异常,且各组间无差别。普通光镜下各组动物肝脏病理切片染色均未见明显异常。结论:目的细胞系细胞培养上清、细胞裂解物对实验用昆明小鼠无明确毒副作用及短期致瘤性,可能提供一种安全的可用于生物人工肝新的细胞来源。
OBJECTIVE: To evaluate acute toxicity and recent tumorigenicity of mouse culture supernatants and cell lysates of HepG2 cell line expressing human liver regeneration enhancing factor gene. Methods: Eighteen SPF Kunming mice were randomly divided into blank control group, cell culture supernatant group, and cell lysate group. Each group of 6 mice was inoculated with blank medium, cell culture supernatant, and cells. Lysate 0.5 ml. For 14 consecutive days, the toxicity of the animals was observed and recorded. After 14 days, the mice were slaughtered. Blood samples were taken for biochemical tests and pathological changes were observed. Results: All mice survived. Except for 1 mouse in the control group, 1 mouse in the cell culture supernatant group, and 2 mice in the cell lysate group had slightly decreased activities on the following day, no abnormal reaction was observed. Blood biochemical tests showed no significant abnormalities in ALT, AST, AFP, and TBIL, and there was no difference between the groups. Under normal light microscope, there was no obvious abnormality in liver pathological staining of each group of animals. Conclusion: The cell culture supernatant and cell lysate of the target cell line have no definite toxicity and short-term tumorigenicity to experimental Kunming mice, and may provide a safe new source of cells for bioartificial liver.