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Study of size selectivity measurement of cell subpopulations using field-flow fractionation
Prokofeva, Daria ; Horstkotte, Burkhard (advisor) ; Chocholouš, Petr (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Analytical Chemistry Candidate: Daria Prokofeva Supervisors: Dr. Burkhard Horstkotte, Ph.D., M.Sc. Consultant: Doc. PharmDr. Hana Sklenářová, Ph.D. Title of Diploma Thesis: Study of size selectivity measurement of cell subpopulation using field-flow fractionation. Cell separation effectiveness of Sedimentation Field Flow Fractionation (SdFFF) was studied by cell isolation and further characterization. In this thesis, we considered only Sedimentation Field-Flow Fractionation (SdFFF). Colorectal cancer (CRC) cell lines - WiDr, HCT116 were used as analytes. Polydispersed complex cell population as CRC cells, contents the cells in different stages of differentiation. It includes the cancer stem cells, which are considered responsible for driving growth and therapy efficacy of treatment. In this study, SdFFF in the so-denoted hyperlayer elution mode was used to obtain cell subpopulations from CRC cell lines. By this, the effectiveness of size-dependent separation and characterization method for cell sorting from complex polydispersed population have been studied. The cell subpopulations (fractions) were collected from different positions of the elution peak. Afterwards, the collected fractions were re-injected to observe the elution...
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