| Objective: To isolate enrich and identify the breast cancer stem cells in human breastcancer MCF-7cell line．Methods: MCF-7cells were suspension cultured in vitro with a serum-free medium(SFM)。the first5days,10days,15days taking pictures, observe sphere-forming cellsis generated. The suspension sphere-forming MCF-7cells were the experimental groupsand the single-layer adherent MCF-7cells cultured with SSM conventionally were thecontrol groups．The percentages of side population（SP）cells in the two kinds of cellswere detected with flow cytometry（FCM）．Results: The MCF-7sphere-forming cells could survive in SFM and form floating cellspheres successfully. The typical loating cell spheres could be observed after5days（0．6±0．5）％and the proportion of cell spheres formed was （2．4±1．2）％inday10． Which was significantly higher than that of day5. The proportion of cellspheres formed wa（s3．8±1．8）in day15．SP cells in theMCF-7floating sphere-formingcells were higher significantly than that of the MCF-7adherent cells（3.9±1.4％vs1.1±0.5％）（P＜0.05）Conclusion: The MCF-7sphere-forming cells which were suspension cultured possesssomproperties of cancer stem cells． Using our suspension culture method with SFMthe breast cancer stem cells in MCF-7cell line can be isolated and enrichedconveniently and effectively．... |
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