技术方法乳腺癌干细胞的培养及鉴定.pdfVIP

2011;31(12) 南方医科大学学报（J South Med Univ ） ·2021 · ·技术方法· 乳腺癌干细胞的培养及鉴定 郑文博，潘凌霄，唐 炜，高 进，叶熹罡 广州医学院第一附属医院乳腺外科，广东 广州 510120 摘要：目的从乳腺癌患者的肿瘤组织中获得乳腺癌干细胞，并确定其生物学特性。方法收集乳腺癌患者的肿瘤组织，利 用乳腺球悬浮培养法获得乳腺球细胞。用流式细胞仪检测细胞CD44 以及CD24 的表达，用Western 印迹法检测细胞 4 5 6 ALDH 1，ESA 以及Oct4 的表达。将2×10 、2×10 和2×10 三种细胞数的原代乳腺球囊细胞及贴壁细胞分别接种到NOD/ + SCID 鼠的脂肪垫，观察其成瘤能力和肝肺转移情况。结果利用乳腺球悬浮培养的原代乳腺癌细胞中有62.36%呈CD44 / CD24-/low 表型，且肿瘤干细胞标记物ALDH 1，ESA 以及Oct4 的表达高于贴壁培养的原代乳腺癌细胞，差异有统计学意义 （P0.05）。与贴壁细胞相比，悬浮培养富集的原代球囊细胞均有更强的成瘤和肝肺转移能力。结论利用乳腺球悬浮培 养法可以从人乳腺癌组织中获得乳腺癌干细胞。 关键词：乳腺癌；肿瘤干细胞；乳腺球悬浮培养 中图分类号：R655.8；R737.9 文献标志码：A 文章编号：1673-4254（2011）12-2021-05 Culture and identification of breast cancer stem cells ZHENG Wen-bo, PAN Ling-xiao, TANG Wei, GAO Jin, YE Xi-gang Department of Breast Surgery, First Affiliated Hospital of Guangzhou Medical College, Guangzhou 510120, China Abstract: Objective To isolate breast cancer stem cells from breast cancer patients and identify their biological characteristics. Methods Mammospheric cells were purified and enriched from the tumor tissues of breast cancer patients using mammosphere culture. Their expressions of CD44 and CD24 were analyzed by flow cytometry, and ALDH1, ESA and Oct4 expressions were determined by Western Blotting. The primary mammospheric and adherent 4 5 6 cells, at the density of 2 ×10 , 2 ×10 or 2 ×10 , were inoculated into NOD/SCID mice to observe their tumorigenic and