New high-speed cell sorting methods for stem cell sorting and breast cancer cell purging

James F. Leary; Scott R. McLaughlin; James A. Hokanson; Judah I. Rosenblatt

doi:10.1117/12.307317

10 April 1998 New high-speed cell sorting methods for stem cell sorting and breast cancer cell purging

James F. Leary, Scott R. McLaughlin, James A. Hokanson, Judah I. Rosenblatt

Proceedings Volume 3259, Systems and Technologies for Clinical Diagnostics and Drug Discovery; (1998) https://doi.org/10.1117/12.307317
Event: BiOS '98 International Biomedical Optics Symposium, 1998, San Jose, CA, United States

Abstract

An important problem in clinical medicine is that of positively selecting hematopoietic stem cells or mobilized peripheral blood stem cells for autologous bone marrow transplantation while purging it of contaminating tumor cells. Since both the stem cells to be positively selected and the tumor cells to be purged are relatively rare cells, this poses special problems for their isolation in terms of purity and yield of stem cells, with a high penalty of misclassification for contaminating tumor cells. A model system of tumor cells spiked into bone marrow or blood cells was used to validate the system. Multiparameter data mixtures of human MCF-7 breast cancer cells and human peripheral blood or bone marrow cells were first analyzed by discriminant function analysis. Mathematical methods were developed to assess the relative probabilities of misclassification. Cell identification tags, implemented as additional correlated listmode parameters not used for these analyses, were used to uniquely identify each cell type and to compare classifier results. The performance of classifier systems was also assessed using ROC (`receiver operating characteristics') analysis. Then the classification system was implemented using lookup tables allowing for real-time (in this system approximately 625 microseconds) rapid separation of these cell types. Isolated cell types, purities and yields were assessed by single-cell PCR molecular characterizations.

Citation Download Citation

James F. Leary, Scott R. McLaughlin, James A. Hokanson, and Judah I. Rosenblatt "New high-speed cell sorting methods for stem cell sorting and breast cancer cell purging", Proc. SPIE 3259, Systems and Technologies for Clinical Diagnostics and Drug Discovery, (10 April 1998); https://doi.org/10.1117/12.307317

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