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. Author manuscript; available in PMC: 2014 May 27.
Published in final edited form as: Nat Protoc. 2012 Jun 7;7(7):1247–1259. doi: 10.1038/nprot.2012.051

Figure 10.

Figure 10

Examples of cell-cell 3D interaction in hydrogel. (a) Co-culture of SMCs (10T 1/2; right channel) and ECs (hMVEC; center channel). Active 3D sprouting angiogenesis into type 1 collagen is monitored (into left hydrogels), while the hMVEC monolayer is stabilized by the presence of SMCs (right hydrogels). 10T 1/2 migration was also activated by the existence of hMVEC monolayer in the center channel. Actin filaments and nuclei were stained with rhodamine-phalloidin (yellow) and DAPI (blue), respectively. (b) GFP transfected U87MG (white arrowheads) transmigrated through the hMVEC monolayer. Colors indicate actin (white), nucleus (blue) and GFP (green). Scale bars 50 μm. (c) Interaction between SMC (white arrowhead) and hMVEC (blue arrowheads). A SMC touches the hMVEC monolayer with its filopodia, and becomes adhered to the monolayer. Colors indicate actin (white), nucleus (blue) and EC-specific primary antibody (green). Scale bars 50 μm. (d) GFP transfected breast cancer cell MDA-MB-231 (white arrowhead) interacting with hMVEC (blue arrowheads) sprouts in type 1 collagen scaffold. Colors indicate actin (white), nucleus (blue) and GFP (green). Scale bars 100 μm. (e) Primary neurons generating axons towards GFP transfected glioblastoma cells U87MG (white arrowheads). Colors indicate actin (white), nucleus (blue) and GFP (green). Scale bars 100 μm.