摘要
背景:肿瘤干细胞与肿瘤的发生和复发存在密切的联系,但是否所有肿瘤中都存在肿瘤干细胞,仍存有一定的争议。目的:探讨人原代胃癌细胞中肿瘤干细胞的分化培养效果。方法:获得新鲜胃癌组织进行原代培养,并进行苏木精-伊红染色和CEA免疫组化鉴定。利用无血清微球体培养体系获得胃癌微球体细胞,免疫荧光法检测微球体细胞CD44表达。以CD44作为标志物对胃癌微球体细胞进行免疫磁珠分选,将分选出的细胞接种于小鼠腋窝后方皮下,观察移植肿瘤生长情况。结果与结论:(1)以无血清培养体系获得人原代胃癌微球体细胞。球体培养获得的细胞中CD44+细胞数量显著多于常规培养获得的细胞(P<0.05);(2)球体培养细胞接种小鼠的移植肿瘤体积增长速度明显快于常规培养细胞接种;(3)移植90 d,获得肿瘤标本,可见球体培养细胞接种小鼠的移植瘤体积明显大于常规培养细胞接种;(4)实验结果表明,利用无血清微球体培养体系结合免疫磁珠分离法可以从胃癌组织中分离出肿瘤干细胞,该细胞具有一定的成瘤性。
BACKGROUND： There is a close relationship between tumor stem cells and tumor occurrence and recurrence, but there are still some disputes in the presence of tumor stem cells in all tumors. OBJECTIVE： To investigate the differentiation and culture of tumor stem cells in human primary gastric cancer cells.METHODS： Primary gastric cancer cells isolated from fresh gastric cancer tissues were stained with hematoxylin-eosin and identified by immunohistochemical detection of carcinoembryonic antigen. The CD44 expression of the cells was detected using immunofluorescence method. Magnetic activated cell sorting was used to isolate CD44＋ gastric cancer cells that were then seeded subcutaneously behind the armpit of mice. Growth of implanted tumor cells was observed. RESULTS AND CONCLUSION： Human primary gastric cancer cells were isolated in serum-free medium. Compared with the routine culture group, the number of CD44＋ cells（P〈0.05） and the tumor volume were significantly increased in the spheroid culture group. Furthermore, at 90 days after transplantation, the tumor volume of mice in spheroid culture group was significantly higher than that in the routine culture group. These experimental findings indicate that gastric cancer cells with certain tumorigenicity can be successfully isolated from gastric cancer cells using serum-free culture method and magnetic activated cell sorting method.
出处
《中国组织工程研究》
CAS
北大核心
2016年第23期3380-3385,共6页
Chinese Journal of Tissue Engineering Research
