Data on Akron Biotech AK Polyfiber Culture Plates
The Florida company of Akron Biotech has been marketing tissue culture dishes and dish inserts which contain their novel and patented polyfiber material. These are polystyrene fibrils which mimic the extracellular matrix and so more accurately model cellular environments in vitro. We obtained two Akron PolyFiber 24 well plates, one which had aligned nanofibrils and the other in which the fibrils were randomly organized. We grew several different cell lines on this, specifically HeLa, Hek293, 3T3 and MDA cells. All cells grew well on all kinds of fiber, and in a few days had almost reached confluence. At this stage we fixed the cells and stained them initially with our mouse monoclonal antibody to actin MCA-5J11 and chicken polyclonal antibody to vimentin CPCA-Vim, using our standard protocol for cells in tissue culture (see here). Both proteins are known to be expressed in all four cell types, and the actin antibody should reveal membranes, ruffles, filopodia and stress fibers while the vimentin antibody should reveal the intermediate filament network in the cytoplasm. The cells were imaged from below the dish using our Olympus FV1000 confocal system fitted to an Olympus inverted microscope. As you can see, we were able to obtain reasonable quality images with no problem by focusing up into the bottom of the dishes, and we have no doubt that it would be possible to image live cells in these dishes if they were labeled with GFP or other in vivo fluorescent compounds. You can also see that cells grown on the random nanofibers have a random orientation, while those grown on the aligned fibers are strongly aligned. So we can say that this product clearly works as advertized.
Legend: Confocal image of HeLa cells grown on Akron's aligned nanofiber network and stained with our mouse monoclonal antibody to actin MCA-5J11 (green) and our chicken polyclonal antibody to vimentin CPCA-Vim (red). Note how the cells are clearly all aligned in a top to bottom orientation, parallel to that of the nanofibers, which can be seen the red channel. Blue staining is the nuclear DNA.
Legend: Confocal image of HeLa cells grown on Akron's random nanofiber network and stained with our mouse monoclonal antibody to actin MCA-5J11 (green) and our chicken polyclonal antibody to vimentin CPCA-Vim (red). Note how the cells are clearly randomly aligned in contrast to the cells shown above. As above the fibers can be weakly visualized in the red channel. Blue staining is the nuclear DNA.
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