【摘要】 目的探讨优化CD45免疫磁珠阴性筛选法对循环肿瘤细胞(CTC)的富集效率。方法利用SW620细胞和健康供体血液样本模拟CTC,加入SW620细胞数分别为50、75、100个,基于细胞角蛋白和上皮细胞黏附分子双抗体染色的CD45阴性筛选法进行CTC富集。比较常规方案正常剂量与高剂量CD45混合物+免疫磁珠富集SW620细胞的效率,以验证CD45混合物和免疫磁珠是否可以通过结合更多的白细胞和包绕更多的CD45-肿瘤细胞来降低肿瘤细胞富集效率;进一步比较优化方案(对含有贴壁CD45+白细胞的试管再次洗脱和磁场分选)与常规方案富集SW620细胞的效率。结果在常规方案中,正常剂量CD45混合物+免疫磁珠富集SW620细胞的平均效率为65.5%,高于高剂量CD45混合物+免疫磁珠的58.3%。采用优化方案富集SW620细胞的平均效率为82.8%,高于常规方案的61.9%。结论优化CD45免疫磁珠阴性筛选法能提高CTC富集效率。更多还原

【Abstract】 Objective To improve the enrichment efficiency of circulating tumor cells(CTC) by optimizing CD45 immunomagnetic bead negative screening method. Methods The testing samples of CTC were simulated by adding 50, 75 and 100 SW620 colon cancer cells in 1 ml healthy donor blood, respectively; and CD45 negative screening method based on CK and Ep CAM double antibody staining was used to enrich CTC. The enrichment efficiency of SW620 cells was compared between normal dose and high dose of CD45 mixture plus immunomagnetic beads to verify whether CD45 mixture and immunomagnetic beads could reduce the enrichment efficiency of tumor cells by binding more white blood cells and wrapping more CD45-tumor cells. The efficiency of the enrichment of SW620 cells between optimized scheme(re-elution and magnetic field sorting of the tube containing adherent CD45+leukocytes) and conventional scheme was compared. Results In the conventional regimen, the average efficiency of CTCs by CD45 mixture plus immunomagnetic beads was 65.5%, which was higher than that of high dose CD45 mixture plus immunomagnetic beads(58.3%). The average efficiency of CTCs by optimized scheme was 82.8%, which was higher than that of conventional scheme(61.9%). Conclusion The optimizated CD45 immunomagnetic bead negative screening method can improve the efficiency of CTC enrichment.更多还原