Ecm select array kit

Non-diseased and malignant endothelial cells were trypsinized, suspended in their respective growth media, and 150,000 cells were seeded onto the ECM Select Array Kit (Advanced Biomatrix #5171) which contained thirty extracellular matrix compositions deposited in quadruplicate onto a hydrogel surface as printed array spots (Kuschel Biotechniques 2006). The arrays were washed 2x at 30 minutes post-seeding in Hank's Balanced Salt Solution (ThermoFisher #14025092) and fixed in 4% paraformaldehyde. Images of each array spot were captured using a digital microscope and cells were counted for each spot. Data was input into a tab- delimited Microscope Excel file and imported into Cluster 3.0 software (http://bonsai.hgc.jp/∼mdehoon/software/cluster/software.htm) where data was centered, normalized, and centroid linkage clustering was performed with a correlation similarity metric. *.CDT files generated in Cluster 3.0 were imported into Java TreeView (http://jtreeview.sourceforge.net/) and heatmaps were generated to represent the quadruplicate attachment preferences of the cell lines for the extracellular matrix compositions on the ECM array.

NALM6 cell line was plated at cell density of 1 × 10⁶ cells/mL onto the ECM Select Array Kit (Advanced Biomatrix) containing 36 different single ECM protein or combinations, each in 9 micro-spots. Twenty-four hours incubation, the ECM Select Array Kit was washed with PBS to remove non-specifically bound cells and was imaged at 4x magnification using an Eclipse TS100 light microscope (Nikon, Melville, NY).