Presentation on theme: "Status Report: Characterization of a panel of cell lines to be used in in the framework of a Treatment Planning System Caterina Tanzarella Antonio Antoccia."— Presentation transcript:
Status Report: Characterization of a panel of cell lines to be used in in the framework of a Treatment Planning System Caterina Tanzarella Antonio Antoccia Antonella Sgura Francesco Berardinelli Dipartimento di Biologia, Università Roma Tre INFN – Sezione di Roma Tre
b. First priority measures in collaboration with the LNL group ENDPOINTS TO BE ANALYZED 1) -H2AX assay for quantitation of tracks at t=30 min 2) Cytogenetic analysis to evaluate the clastogen effect in IR-exposed cells. c. Additional measures 2) Cell inactivation to evaluate the surviving fraction in IR-exposed cells a.Characterization of cell lines 1) Nuclear area 2) Nuclear thickness In V79, AG01522D, CCD18Lu, HSG, T98G 1) Cell growth curves
Measure of nuclear area -Cells were seeded at a final concentration of 12000/cm 2 in chamber slides successively fixed in absolute methanol and stained with 0,5µg/ml DAPI - Images were captured at 63X magnification and nuclear area evaluated using the ISIS sofware by Metasystems
~ 0,38 µm Thickness (µm) = (number of slices -1) * thickness of a single slice (µm) BF Each cell was analysed indipendently and at least 100 cells were scored for each cell line Measure of nuclear thickness Analysis performed in vivo using DRAQ5 dye in confocal microscopy
Nuclear area and thickness in AG01522 D
Nuclear area and thickness in V79
Cell growth of AG01522 D Doubling time: ~ 22 hrs T = t log2 / (log N -log N0) Cells were seeded at a final concentration of 16*10 4 in plastic Petri dishes and counted in triplicate every 24 hours for 5 days.