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目的人乳腺癌肿瘤移植裸鼠模型的构建及其肿瘤浸润髓系细胞的分离。方法将p HAGE-EF-Zs GDEST质粒与病毒包装质粒p MD2.G和ps PAX2首先通过磷酸钙转染方式包装带绿色荧光的慢病毒,再利用包装好的慢病毒感染BT474细胞,通过流式分选筛选阳性细胞,扩大培养后,种植于裸鼠乳腺脂肪垫,应用活体荧光成像仪和体外测量肿瘤大小,对人-裸鼠肿瘤移植模型的构建效果进行评价;通过机械法分离该小鼠模型的肿瘤浸润髓系细胞。结果筛选出带绿色荧光的BT474稳定株,种植于裸鼠乳腺脂肪垫,构建了人-裸鼠肿瘤移植模型,并分离出其肿瘤浸润髓系细胞。结论成功分离出了肿瘤浸润髓系细胞,为后续研究肿瘤浸润髓系细胞的功能奠定了基础。
OBJECTIVE: To construct the nude mice model of human breast cancer xenograft and to isolate the tumor-infiltrating myeloid cells. Methods Plasmid p HAGE-EF-Zs GDEST and viral packaging plasmid p MD2.G and ps PAX2 were first packaged with green fluorescent lentivirus by calcium phosphate transfection and infected with BT474 cells by packaged lentivirus. The positive cells were screened and selected for expansion. After being cultured, the mice were inoculated with mammary fat pads in nude mice. The in vivo fluorescence imaging was used to measure the size of the tumors. The effect of constructing the tumor-bearing model in human-nude mice was evaluated. Model tumors infiltrate myeloid cells. Results The stable green fluorescent BT474 strain was screened and implanted into the mammary fat pad of nude mice. The human and mouse tumor model was established and the myeloid cells of tumor infiltrated were isolated. Conclusions The tumor infiltrating myeloid cells was successfully isolated, which lays the foundation for further study on the function of tumor infiltrating myeloid cells.