人结肠癌CW-2干细胞获取方法研究

目的运用3种不同的方法分离纯化人结肠癌CW-2干细胞，并对其分离纯化效率进行比较，探讨获得癌干细胞的有效方法。方法采用单纯无血清悬浮培养、无血清悬浮培养联合化疗药物、流式细胞分选技术分别富集人结肠癌细胞株CW-2干细胞；然后运用流式细胞术、NOD—SCID小鼠致瘤实验和Transwell侵袭实验分析比较3种方法的富集效率。结果无血清悬浮培养细胞．无血清悬浮培养联合化疗药物处理细胞和流式细胞仪分选技术分选后细胞中具有结肠癌干细胞特性的CD44＋EPCAM_细胞分别为（59．39±4．55）％、（74.36±6．78）％、（86．43±8．43）％；3群细胞的成瘤能力和侵袭能力都存在显著统计学差异（P值〈0．05）：流式细胞分选技术分选后细胞〉无血清悬浮培养联合化疗药物处理细胞〉单纯无血清悬浮培养细胞。结论流式细胞分选技术富集癌干细胞的能力强于单纯无血清悬浮培养和无血清悬浮培养联合化疗药物，无血清悬浮培养联合化疗药物又强于单纯无血清悬浮培养。

Comparison of Methods of the Isolation Colon Cancer Stem Cells from Human Colon Cancer Cell Line CW-2

Objective Using three different ways to isolate colon cancer stem cells from human colon cancer cell line CW-2, to evaluate the purification efficiency and find out the most efficient way. Methods CW-2 stem cells were enriched with three methods which are simple ser-um-free medium,serum-free medium combination with anticancer drugs, FACS (Fluorescence-activated cell sorting) technique; and then compared the enrichment efficiency of the three methods with flow cytometry,fumorigericity assay in NOD-SCID mice and Transwell inrasier assay. Results CD 44 + EPCAM + cells characterized of colon cancer stem cells were (59.39±4.55)%, (74.36±6.78 ) %, (86.43±8.43)% separately in cells enriched by in simple serum-free medium, serum-free medium combination with anticancer drug and FASS;tumor formation capability and invasive capacity of the three group cells had statistically significant differences(P<0.05):cells enriched by FACS were greater than cells cultured in serum-free medium combination with anticancer drug,cells cultured in that were greater than cells cultured in simple serum-free medium. Conclusion Enrichment capability of cancer stem cells of Facs techhique was higher than in pure serum-free medium and serum-free medium anticancer with anficancer drug,the enrichment capability of serum-free medium anficancer with chemothera-py drug was higher than pure serum-free medium.