Separation of a Breast Cancer Cell Line from Human Blood Using a Quadrupole Magnetic Flow Sorter

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• 作者：Masayuki Nakamura ; Keith Decker ; Julia Chosy ; Kristin Comella ; Kristie Melnik ; Lee Moore ; Larry C. Lasky ; Maciej Zborowski ; and Jeffrey J. Chalmers
• 刊名：Biotechnology Progress
• 出版年：2001
• 出版时间：December 2001
• 年：2001
• 卷：17
• 期：6
• 页码：1145 - 1155
• 全文大小：189K
• 年卷期：v.17,no.6(December 2001)
• ISSN：1520-6033

文摘

We have developed a quadrupole magnetic flow sorter (QMS) to facilitate high-throughput binary cell separation. Optimized QMS operation requires the adjustmentof three flow parameters based on the immunomagnetic characteristics of the targetcell sample. To overcome the inefficiency of semiempirical operation/optimization ofQMS flow parameters, a theoretical model of the QMS sorting process was developed.Application of this model requires measurement of the magnetophoretic mobilitydistribution of the cell sample by the cell tracking velocimetry (CTV) techniquedeveloped in our laboratory. In this work, the theoretical model was experimentallytested using breast carcinoma cells (HCC1954) overexpressing the HER-2/neu gene,and peripheral blood leukocytes (PBLs). The magnetophoretic mobility distributionof immunomagnetically labeled HCC1954 cells was measured using the CTV technique,and then theoretical predictions of sorting recoveries were calculated. Mean magnetophoretic mobilities of (1-3) × 10^-4 mm³/(T A s) were obtained depending on thelabeling conditions. Labeled HCC1954 cells were mixed with unlabeled PBLs to forma "spiked" sample to be separated by the QMS. Fractional recoveries of cells fordifferent flow parameters were examined and compared with theoretical predictions.Experimental results showed that the theoretical model accurately predicted fractionalrecoveries of HCC1954 cells. High-throughput (3.29 × 10⁵ cells/s) separations withhigh recovery (0.89) of HCC1954 cells were achieved.