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. Author manuscript; available in PMC: 2013 Dec 2.
Published in final edited form as: Methods Enzymol. 2012;506:10.1016/B978-0-12-391856-7.00034-2. doi: 10.1016/B978-0-12-391856-7.00034-2

Figure 10.2.

Figure 10.2

MAME tripartite cocultures of human SUM102 breast carcinoma cells (red; transduced with Lenti-RFP) with human umbilicial vein endothelial cells (HUVEC; blue; stained with CellTrace Far Red) in reconstituted basement membrane above a bottom layer of WS-12Ti human breast tumor-associated fibroblasts (magenta; transduced with Lenti-YFP) in interstitial collagen I (see also Fig. 10.3A). At 2 days of coculture as illustrated here, the SUM102 cells cluster around the branching networks formed by the HUVECs. Degradation products of DQ-collagen IV (green) are apparent around the interacting cells. The fibroblasts, which are primarily in the layer of collagen I, are associated with pericellular degradation products of DQ-collagen I (white). A few fibroblasts, also associated with proteolysis, can be seen to have migrated into the upper layer. The three panels from left to right are 3D reconstructions of the MAME tripartite coculture from the top, the bottom and at a 45° angle. Magnification, 20×.