Human beta cell cultures growing in vitro. Morphological and immunofluorescence studies performed on cells from passages 5 to 15. (A) Cells (1 × 104 cells/well) obtained from human insulin secreting cell cultures, were incubated for 4 days in CMRL medium containing 0.5% FCS. 2-dimensional, expansive growth pattern (top panel) and a cluster-like, 3-dimensional type of growth (bottom panel). Representative fields (x100) were photographed under a phase-contrast Nikon microscope. (B) Representative G-banded karyotype of VGA (46, XX) (upper left panel); APM (46, XY) (upper right panel) and CPR cells (46, XY) (bottom left panel) and (46, XY, der(22)) (bottom right panel, black arrow indicates chromosome 22 alteration). (C) Immunoflourescence confocal microscopy. After fixation and permeabilization, the cells were labeled with antibodies against chromogranin A(green) and alpha-actin (red) (top left panels), insulin (green) and desmin (red) (middle left panels), glucagon (green) and desmin (red) (bottom lef panels), Glut2 (green) and vimentin (red) (top right panels), C-peptide (green) and vimentin (red) (middle right panels) or PDX1 (green) and cytokeratin 19 (red) (bottom right panels). In all cases nuclei were stained with DAPI (blue). A representative field is shown. Bar: 50 μm.
Labriola et al. BMC Cell Biology 2009 10:49 doi:10.1186/1471-2121-10-49