Fig. 3
a. Change in the morphology of CAM by the invasion of SaOS2 cells. Images of vertical cross sections of CAM stained with haematoxylin and eosin after incubated with 20 μl volumes of 3 × 105, 6 × 105 and 12 × 105 SaOS2 cells along with DMEM alone as negative control for 96 h. CAM incubated with all three concentrations of SaOS2 cells shows increased fibroblast accumulation and irregular growth pattern of thin stratum at ectodermal layer and stroma shows presence of numerous small blood vessels around the larger ones (b, c and d). Ruptured ectodermal layer with invading tumor growth is prominent with all three concentrations and the invading area shows the presence of blood vessels (arrow head) with nucleated erythrocytes (pink color, arrow) and the presence of numerous monocytes (b’, c’ and d’). Control CAM shows uniform thickness at the ectodermal and more fibroblast proliferation (a) and the presence of nucleated erythrocytes (in pink colour) trapped in the normal blood vessels (a’). n=3 and bar is 50 μm. Figures a’, b’, c’ and d’ are enlarged version of the squared area of the same. ibv- large blood vessel, sbv- small blood vessel, v- vein, ce and ae-chorionic and allantoic epithelial layers and SEC- sub epithelial capillary network. Arrow shows the presence of tumor invasion and growth in to the ectodermal layer of CAM in b, c and d. b. SaOS2 cells induces the growth and thickness of CAM. Distances between chorionic and allantoic layers was measured morphometrically in micrometer excluding the tumor growth from haematoxylin and eosin stained cross section of CAM after incubation. CAM incubated with 6 × 105 and 12 × 105 SaOS2 cells shows significant increase in the tissue thickness than control. Each value is the mean ± SEM, p = < 0.001 versus control, n = 6