Magnetic activated cell sorting combined with a panel of monoclonal antibodies in detecting free cancer cells in analogue malignant ascites / 中华消化杂志

ABSTRACT

Background Magnetic activated cell sorting(MACS)is a new immunomagnetic separation technique.It's principle is based on the specialized recognition between the antibody and antigen.When directed or indirected coupled with the 50 nm magnetic beads,the antibody can linked with the cells which express the corresponding antigen,and then leads to the magnetic cell separation in a high intensity and high-gradient magnetic field.The method has higher separative purity and recovery and been made use of enriching tumor cells and tumor stem cells.It could be also utilized to enrich the rare tumor cells in fluid specimen.Objective To evaluate the enriching efficiency of detecting free cancer cells in analogue malignant ascites using MACS and a panel of tumor-specific markers.Methods Five species of tumor cell lines correlated to the diseases resulting in malignant ascites were selected and the expressions of EpCAM,CAl25,CEA,TAG-72 and their mixture in these tumor cell lines were detected using immunofluorescence reactions and flow cytometry(FCM).The tumor cells were spiked into mononuclear cells by different ratios to analogue the main ingredients of malignant ascites.The efficiency of MACS combinded with mixture antibodies in separating tumor cells was compared with single antibody.Results The FCM revealed that the expression of the mixture antibodies in 5 tumor cancer cell lines was significantly higher than that in single antibody.The mean recovery rates of spiked tumor cells were enhanced by using a panel of monoclonal antibodies combined with MACS than single antibody,especially in two gastric cancer cells and colon cancer cells(69.18%±20.84%VS 45.23%±11.54%,78.75%±15.42%vs 59.73%±16.64%and 85.63%±12.30%VS 76.88%±8.65%,respectively),the ovarian cancer cells was the next(32.49%±3.58%vs31.79%±4.82%),and the liver cancer cells was the lowest(11.78%±0.43%VS 7.16%±0.46%).Conclusions The detection of free cancer cells from malignant ascites by MACS combined with a panel of monoelonal antibodies is more effectively than single antibody.The method has the potencial value of clinical application to the malignant ascites caused by gastroenteric tumor.