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目的按照常用肿瘤细胞建系方法,以人卵巢上皮癌组织移植免疫缺陷小鼠,通过原代培养建立永生化细胞系,为人体肿瘤的研究提供体外研究模型。方法将1例卵巢浆液性乳头状癌IIIcG3患者手术切除的肿瘤组织接种裸鼠皮下成瘤,通过体外原代培养,建立1株悬浮生长的细胞系。通过光学显微镜、电子显微镜、生长曲线测定、染色体分析、克隆形成实验、双层软琼脂培养、裸鼠接种等,对其生物学特性进行研究。结果该细胞系已传至100代以上,命名为WSZ。其生物学特性为:形态学观察细胞呈悬浮球形生长状态;细胞生长增殖旺盛,对数期细胞群体倍增时间约15.8h;染色体为16～135条,以68和69条染色体为多见,染色体分析显示,基本全部为端着丝粒染色体;细胞克隆形成率为89.3%,具有软琼脂集落形成能力;异种移植实验表明106细胞在裸鼠皮下可成瘤,而100个细胞接种非肥胖型糖尿病/重症联合免疫缺陷(NOD/SCID)小鼠即可形成皮下肿瘤。结论 WSZ能够在体外长期生长和稳定传代,WSZ是一高度恶性的细胞系。
Objective To construct immortalized cell lines by primary culture in order to provide immunosuppressed mice with epithelial ovarian cancer and to provide in vitro models for the study of human tumors. Methods One tumor of ovarian serous papillary carcinoma IIIcG3 was inoculated subcutaneously into nude mice. Primary suspension culture was used to establish a cell line that grew in suspension. The biological characteristics were studied by optical microscope, electron microscope, growth curve determination, chromosome analysis, clone formation experiment, double soft agar culture and nude mice inoculation. Results The cell line has been transmitted to more than 100 generations, named WSZ. The biological characteristics of the cells were as follows: morphological observation showed that the cells were in the form of suspended spherical growth; cell growth and proliferation were strong, the population doubling time was about 15.8 hours in logarithmic phase; chromosomes were 16-135, with 68 and 69 chromosomes being common, chromosomes The analysis showed that all of them were telocentric chromosomes. The rate of cell colony formation was 89.3%, with soft agar colony forming ability. The xenograft experiments showed that 106 cells could be subcutaneously implanted in nude mice, while 100 cells were inoculated with non-obese diabetic / Severe combined immunodeficiency (NOD / SCID) mice to form subcutaneous tumors. Conclusion WSZ can grow long-term in vitro and stable passage, WSZ is a highly malignant cell line.