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目的利用肝门部胆管癌原发病灶建立的细胞系接种裸鼠脾脏,筛选出高转移亚系。方法将FRH-0201制成细胞悬液,接种7只BALB/C裸鼠脾脏。将转移的肿瘤行组织块培养,筛选出细胞亚系。FRH-A再次接种裸鼠,将转移瘤再次培养,筛选出细胞亚系FRH-B。观察细胞形态学、核型变化、肿瘤标志物、角蛋白染色及细胞生长曲线。结果FRH-A及FRH-B细胞系为梭形和圆形细胞,生长旺盛。电镜显示典型恶性细胞特征。细胞倍增时间为21.8 h,染色体众数19条,主流范围18~44条。细胞上清液CA125显著增高。细胞角蛋白染色阳性。FRH-A细胞再次接种裸小鼠,转移发生率100%,肿瘤生长迅速,转移发生快。结论FRH-A及FRH-B细胞系来源于FRH-0201的高转移的亚系。
OBJECTIVE: To clone the spleen of nude mice using the cell line established by the primary lesion of hilar cholangiocarcinoma and screen out the highly metastatic subfamilies. Methods FRH-0201 cells were made into a cell suspension and inoculated 7 BALB / C nude mice spleen. Tumor tissue metastasis will be cultured to screen out cell sublines. The nude mice were inoculated again with FRH-A, the metastases were cultured again, and the cell subline FRH-B was screened out. Cell morphology, karyotype changes, tumor markers, keratin staining and cell growth curve were observed. Results The FRH-A and FRH-B cell lines were spindle and round cells with vigorous growth. Electron microscopy showed typical malignant cell features. Cell doubling time was 21.8 h, chromosome number 19, the mainstream of 18 to 44. Cell supernatant CA125 was significantly higher. Cytokeratin staining positive. FRH-A cells were inoculated nude mice again, the incidence of metastasis 100%, rapid tumor growth, metastasis occurs quickly. Conclusion The FRH-A and FRH-B cell lines are derived from the highly metastatic subline of FRH-0201.