Fig 1.
Primary canine inflammatory mammary carcinoma origin of the cell line IPC-366.
Tumor paraffin sections, H&E. A (10X) y B (20X). Neoplastic emboli in superficial dermis. Tumor cells exhibit marked anisocitosis and anisokaryosis, and evident large nucleoli. Infiltrating tumor cells (arrow).
Table 1.
Technical data of specific antibodies and peroxidase-developing systems for immunohistochemistry.
Fig 2.
A) IPC-366 cells in culture at inverted microscopy (40X). B) IPC-366 pellet; paraffin section, H&E (40X). Highly malignant tumor cells with epithelial morphology; marked anisocytosis and anisokaryosis and evident nucleoli. Endothelial-like cell (ELC) showing cytoplasmic clear space (arrow). IPC-366 ultrastructural features were evaluated by electron microscopy. The cells had abundant cytoplasmic projections, numerous organelles and proteinaceous secretory products. By electron microscopy, the clear cytoplasmic vacuoles seen by optic microscopy, resulted empty spaces lined by cytoplasmic membranes that occasionally joined to form an internal lumen (ELCs, vasculogenic mimicry). (Fig. 3C-H).
Fig 3.
IPC-366 cells in pellet resin sections.
A and B) Semithin sections stained with toluidin blue (40X); some endothelial-like cells are observed (arrows). C to H) Ultraestructure in ultrathin sections. C) Cells with large nuclei and evident nucleoli. Mitosis (arrow). D) Single cell with elongated nucleus, very evident nucleolus and numerous organelles, vacuoles and proteinaceous secretion. E) Tumor degenerated cell with shrunken nucleus and abundant proteinaceous secretion. F) Cytoplasm of a tumor cell containing numerous organelles (mitochondria, endoplasmic reticulum) and a secretory vacuole. G) Tumor cell with three cytoplasmic empty spaces (asterisks) covered by cytoplasmic membrane; progression to a cytoplasmic “lumen” characteristic of endothelial-like cells. Evident nucleolus. H) Binucleate tumor cell with a cytoplasmic central empty space (“lumen”, asterisk) (ELC) resembling a capillary vessel (vasculogenic mimicry). Figs. E and G show different fields from the same grid. At 25th passage, cells were cryogenic storage (-180°C). Re-cultured thawed cells presented similar growth and morphological characteristics with a viability of 95–99%. The doubling time of IPC-366 cells at 30th passage was approximately 24 hours and the cells reached a plateau on day 6 (Fig. 4).
Fig 4.
Growth curve of IPC-366 at 30th passage.
Fig 5.
Tumors from mice inoculated with IPC-366.
A) IPC-366 mouse xenografts at four weeks (arrow). B to E: tumor paraffin sections H&E. B) Solid tumor infiltrating the dermis (10X). C) Neoplastic embolus in a superficial dermal lymphatic vessel (arrow) and solid tumor; marked edema in dermis (4X). D) Highly malignant tumor cells with marked anisocytosis and anisokaryosis; evident nucleoli (40X).E) Microvascular channels lined up by neoplastic cells (vasculogenic mimicry, arrows) (20X).
Fig 6.
Immunohistochemistry of IPC-366 pellet, paraffin sections.
A) Pancytokeratin (AE1/AE3) (10X); intense immunolabeling and numerous positive cells. B) CK14; intense immunolabeling and numerous positive cells (10X). C) Vimentin; moderate immunolabeling and numerous positive cells (10X). D) p63; negative (20X). E) α-smooth muscle actin, negative cells (10X). F) Estrogen receptor; negative cells (20X). G) Progesterone receptor; negative cells (20X). H) HER-2; negative cells (20X). I) E-Cadherin; intense membranous immunolabeling in numerous cells (20X). J) COX-2; most of the cells are moderately positive; some cells are intensely positive (10X). K) COX-2; some cells strongly positive showing features of malignancy (binucleated cell, arrow) (40X). L) Ki-67 proliferation marker; many nuclei are positive (10X).
Table 2.
Aberration found in the Karyotype.