Dispase 1 solution

We followed the method described by Berbéri et al. [17 (link)]. Briefly, hMSSM samples were extensively washed with PBS supplemented with 1% P/S and cut into small pieces under aseptic conditions. Tissue fragments were incubated with 1 U/ml dispase I solution (Sigma-Aldrich, USA) in PBS at 37°C for 1 hour to separate the epithelial lining from the membrane. Epithelial cells were discarded, and the remaining tissue fragments were treated with 200 collagen digestion units (CDU)/ml of collagenase type II (Sigma-Aldrich, USA) in Hank's balanced salt solution (HBSS) containing 5 Mm calcium chloride at 37°C for 3 hours. Tissues were shook repeatedly during enzymatic incubation. The resulting cells were filtered out with a 40 μm cell strainer (BD Biosciences), and then, hMSSM-derived cells were centrifuged at 900 RPM for 10 minutes.

We followed the method described by Berberi et al. 2016 [24 (link)]. hMSSM samples were extensively washed with PBS supplemented with 1% P/S and cut into small pieces under aseptic conditions. Tissue fragments were incubated with 1 U/ml dispase I solution (Sigma-Aldrich, USA) in PBS at 37°C for 1 hour to separate the epithelial lining from the membrane. Epithelial cells were discarded, and the remaining tissue fragments were treated with 200 collagen digestion units (CDU)/ml of collagenase type II (Sigma-Aldrich, USA) in Hank's balanced salt solution (HBSS) containing 5 Mm calcium chloride at 37°C for 3 hours. The tissues were shaken repeatedly during enzymatic incubation. The resulting cells were filtered out with a 40 μm cell strainer (BD Bioscience). The hMSSM-derived cells were then centrifuged at 900 RPM for 10 minutes.