Which extracellular matrix (ECM) do you recommend for culturing your endothelial cells?
We recommend culturing iCell® Endothelial Cells on tissue culture plates coated for 1 hour at room temperature with 3 µg/cm2 fibronectin. Other extracellular matrixes (collagen IV, gelatin, etc.) have been used by customers, and Belair et al (2015) demonstrated angiogenesis endpoints on 2D and 3D Matrigel ECMs. To coat surfaces, dilute stock fibronectin in sterile water to a final concentration of 30 µg/ml, add volume shown in the table below and incubate at room temperature for 1 hour. Note that we recommend using freshly coated vessels, but vessels may be coated stored in fibronectin (wrapped in Parafilm sealant to prevent evaporation) at 4°C for up to 1 week and warmed for 1 hour at room temperature before use. We do not recommend using culture vessels if the fibronectin solution has evaporated. Note that the ECM should be present in culture vessels for all culture steps (thawing, plating, maintenance, and passaging).
|Culture Vessel||Coating Volume|
|T-75 Flask||7.5 ml|