Figure 1

Culture and characterisation of a clonal luciferase-expressing bone marrow-derived stromal cell line from ROSA26-L-S-L-Luciferase transgenic mice. (A) Molecular organisation of the ROSA26 locus in ROSA26-L-S-L-Luciferase transgenic mice. (B) Representative pictures of cultured bone marrow-derived stromal cells (BMSC) taken under phase contrast microscopy. Left: unmodified parental BMSC derived from ROSA26-L-S-L-Luciferase transgenic mice (BMSC parental). Right: clonal luciferase-expressing BMSC derived from ROSA26-L-S-L-Luciferase transgenic mice (BMSC-Luc clonal). (C) Parental BMSC derived from ROSA26-L-S-L-Luciferase transgenic mice were non-electroporated (NO EP, upper dot plot) or electroporated with eGFP mRNA (EP eGFP, lower dot plot), and were analyzed by flow cytometry for eGFP fluorescence (x-axis) versus viability (GelRed-staining, y-axis) after 24 hours of culture. The percentage indicated in the lower left quadrant is the number of viable eGFP-negative cells. The percentage indicated in the lower right quadrant is the number of viable eGFP-positive cells. The percentages indicated in the upper left and right quadrant are numbers of non-viable cells. Representative dot plots are shown. (D) In vitro luminescence assay on parental BMSC (BMSC parental), on Cre-recombined polyclonal luciferase-expressing BMSC (BMSC-Luc polyclonal), and on Cre-recombined clonal luciferase-expressing BMSC (BMSC-Luc clonal), all derived from ROSA26-L-S-L-Luciferase transgenic mice. (E) Representative flow cytometric analysis showing expression pattern of membrane proteins on clonal BMSC-Luc derived from ROSA26-L-S-L-Luciferase transgenic mice (i.e. expression of Sca-1, V-CAM and MHC-I, but no expression of MHC-II, c-kit, CD45, CD31 and A2B5). Open histograms: control. Filled histograms: specific antibody staining.