Cpd 030

Cell morphology and attachment on scaffolds was determined by SEM using an XL 30 ESEM-FEG (Philips, The Netherlands). Briefly, samples were rinsed twice with PBS, fixed with 10% formalin for 2 h. After washing with PBS two times, the samples were dehydrated with a gradient ethanol series, dried by using critical point drying (Balzers CPD 030) and sputtered with gold (Cressington) before SEM analysis.

To study the effect of the mCP ECM proteins on INS1E cell morphology, samples were fixed in 4% (w/v) paraformaldehyde in PBS for 1 h at room temperature after 1, 3, and 7 days of culture. Subsequently, samples were dehydrated using an increasing concentration series of ethanol from 70% to 100% for 30 min in each step. Afterwards, samples were prepared for scanning electron microscopy using a Balzer's CPD 030 and BAL-TEC critical point dryer, and gold coated by a sputter coater (Cressington, UK). Electron micrographs were made using an XL30 ESEM-FEG environmental scanning electron microscope (Philips/FEI, The Netherlands). Furthermore, to verify cell growth, samples from each experimental condition were fixed in 4% (w/v) paraformaldehyde in PBS for 1 h at room temperature during cell culture at 3 and 7 days and imaged using standard bright-field phase contrast microscopy (Nikon TE300, Japan).