Figure 3.

Embedding procedure and slicing. Images showing the steps of tissue preparation for cutting epicardial slices. (a) The ventricles are isolated avoiding any direct handling of the epicardium and placed on ice throughout the procedure. (b) Tissue blocks are dissected by making an incision through the ventricular wall with a single edge steel blade and (c) placed on cold agarose inside a 3D printed plastic ring, the epicardium facing down. (d) The plastic ring is filled with low melting agarose. (e) Once solidified, the block is removed from the ring, squared and mounted on the holder using cyanoacrylate glue. (f) Epicardial slices are cut using a vibratome, with the blade speed at 0.03 mm/s. (g–i) Epicardial slices are pinned to pillared Petri dishes, and either cultured in static conditions changing medium every 24 h, or connected to BioFlo120^Ⓡ benchtop bioprocessor through a custom 3D-printed insert, enabling flow culture and control of the culture conditions. Source: Maselli et al. (23).