免疫磁珠法分离纯化人肾癌CD133~+细胞实验研究

目的:本研究通过分析CD133+细胞在肾细胞癌患者肿瘤组织的表达情况,寻找肾癌中CD133+细胞分离纯化方法,探寻CD133+细胞与肾癌发生中的作用。方法:①应用胎盼蓝拒染试验检测细胞活性。②应用免疫磁珠法分离纯化肿瘤组织中CD133+细胞。③应用流式细胞仪检测肾透明细胞癌标本中CD133+细胞表达情况。结果:30例肾透明细胞癌中CD133+比为(13.08±1.39)%,与病理分级、临床分期无关。免疫磁珠法分离纯化CD133+纯度为(75±2.4)%,细胞活性不受影响。结论:免疫磁珠技术纯化肾癌CD133+细胞成熟有效,CD133+细胞可能是肾癌干细胞。

Primary study to isolate the CD133~+ cells from the human renal cell carcinoma by the immunomagnetic microbeads WU

Objective:To evaluate the role of CD133 + progenitor cells in the occurring of renal cell carcinoma ,we performed cytofluorimetric analysis to renal cell carcinoma,established the method of isolation and purify the CD133 + cells by the immunomagnetic microbeads. Methods: We investigated by cyto fluorimetric analysis the presence of a cell population expressing the CD133 marker in cells dissociated from tissue specimens of 30 renal carcinomas. CD133 + cells were isolated by magnetic cell sorting,using the MACS system. The CD133 + cells and CD133-cells were performed microarray after magnetic isolation . Results: All carcinomas displayed a CD133 + cell population with a mean percentage(13.08±1.39)% of the total cells. No correlation among the number of CD133 + cells and tumor stage or grade or patient age or sex was observed. The purity of CD133 + cells was about (75±2.4)% and the viability of cells was not affected by the MACS,comparing the expression of CD133 + cells with CD133 -cells. Conclusion: The CD133 + progenitor cells is probably the initiating cells of renal cell carcinomal,and the method of MACS is perspective for the use of isolation renal cancer stem cells.