Figure 1.

Study design and rapid IL-10 production of cryopreserved MSCs^IL-10 in vitro. (A) Human umbilical cord MSCs were genetically engineered for augmented human IL-10 production (MSCs^IL-10) and cryopreserved. (B) In vitro bench testing of the cryopreserved MSC^IL-10 batches for hIL-10 production (54 × 10³ cells from the 20 × 10⁶ MSC^IL-10 group, n = 5; or from the 40 × 10⁶ MSC^IL-10 group, n = 3). (C) Pig double lungs subjected to 24 h cold ischemia were connected to EVLP for 6 h and randomized to control (n = 7), or pulmonary artery delivery of 20 × 10⁶ (n = 5) or 40 × 10⁶ cryopreserved MSCs^IL-10 (n = 6). Subsequently, the left lung was transplanted into a recipient pig that received methylprednisolone and cyclosporine A background immunosuppression, and was sacrificed 3 days after transplantation. (D) A clinical-grade Toronto ex vivo lung perfusion system was used for the EVLP experiments, and MSCs^IL-10 were administered to the EVLP circuit in a blinded and randomized manner with a syringe (white arrow) and a tubing extension was attached to the pulmonary artery cannula (black arrow). (E) Analysis of preformed hIL-10 in MSC^IL-10 supernatant taken immediately prior to MSC^IL-10 administration. Data mean ± standard deviation and analyzed with (B) two-way ANOVA using Šidák correction for multiple comparisons or (E) 2-tailed Student’s t-test. Ad, adenovirus; EVLP, ex vivo lung prefusion; hIL-10, human interleukin-10; MSC, mesenchymal stromal cell; TX, transplantation.