Heparin sodium salt from the porcine intestinal mucosa

To induce smooth muscle differentiation of MPC, murine MPC and MSC to MPC-SMC, and murine MPC-SMC and MSC-SMC, cells cultivated with growth medium to a confluency of about 70% were washed once with DMEM/F12 (Thermo Scientific, MA, USA). Next, cells were covered with smooth muscle differentiation medium, consisting of DMEM/F12 supplemented with recombinant human TGFb1 (Thermo Scientific, MA, USA), heparin sodium salt from the porcine intestinal mucosa (Sigma-Aldrich Co. LLC, MO, USA), fetal calf serum (Gibco, Thermo Scientific, MA, USA), and gentamicin (Sandoz GmbH, Tirol, Austria) to a final concentration of 10 ng/ml, 5% (v/v), and 3.84 μg/ml, respectively. Finally, cells were cultivated in smooth muscle differentiation medium for 3–6 days at 37 °C, 5% CO₂. The medium was changed every 3–4 days.