条件性重编程技术分离培养人原代乳腺癌干细胞

目的 应用条件性重编程细胞(Conditionally Reprogrammed Cells,CRCs)技术建立高效、稳定的人乳腺癌千细胞体外培养,建立潜在的乳腺癌干细胞靶向药研究模型.方法 收集人乳腺癌组织样本,采用CRCs技术分离培养原代乳腺癌细胞,用CD24免疫磁珠标记原代乳腺癌细胞后使用磁性细胞分选仪分选出乳腺...

Isolation and Cultivation of Human Primary Breast Cancer Stem Cells Based on Conditionally Reprogrammed Cells Technique

OBJECTIVE To establish efficient and stable human breast cancer stem cells in vitro by conditionally reprogrammed ceus(CRCs)technique,and to establish potential breast cancer stem cells targeted drug research model.METHODS Primary breast cancer cells were isolated from breast cancer patients surgical specimens and cultured by CRCs technique,and breast cancer stem cells were then selected by magnetic cell sorter after primary breast cancer cells being labeled with CD24 immunomagnetic beads.The expression of Pan Cytokeratin(pan-CK)was detected by immunofluorescence.The expression levels of cancer stem cells biomarkers CD44,CD24,and ALDH1 was measured by flow cytometry.The cell clonies,and formation of mammospheres were determined by colony formation experiment,and mammospheres formation experiment.RESULTS The primary breast cancer cells isolated and cultivated by CRCs technique could be continuously cultivated in vitro for up to 9 passages,and the characteristics of breast cancer were maintained.After the primary breast cancer cells were cultured by CRCs technique and sorted by magnetic cell sorter,the mammosphere formation rate and clony formation rate of breast cancer non-stem cells were significantly lower than that of breast cancer stem cells.Breast cancer stem cells could be passaged to 5 passages.CONCLUSION Breast cancer stem cells can be isolated efficiently from primary breast cancer tissues and can be proliferated stably by the combined applicaiton of CRCs and magnetic cell sorter.And the system has a strong potential to be a new breast cancer stem cells targeted drug research model.