Figure 2.

The functionality of MSCs is not compromised with pronase. (A): Pronase-detached umbilical cord blood (UCB)-MSCs were able to inhibit T-cell proliferation as effectively as control cells. T-cell proliferation was determined by measuring the amount of 5(6)-carboxyfluorescein diacetate N-succinimidyl ester (CFSE)^low cells gated from leukocyte population on the forward and side scatter plot. Responder cells (positive proliferation control) were assigned as 100%. Data are represented as mean (+SD) of two UCB-MSC donors against allogeneic peripheral blood mononuclear cells from two different healthy donors (n = 4). (B): The angiogenic ability of pronase-detached UCB-MSCs in the in vitro assay was comparable to that of control cells. In the coculture setup, the angiogenic ability was as high as or even higher than that of positive controls (assigned as 100%), but in inserts remained on the same level of induction as the medium control. Data are represented as mean (+SD) (n = 3). (C): Different rolling speed profiles of trypsin- and pronase-detached BM-MSCs in flow perfusion assay on a fibronectin-coated capillary. Cells having a rolling speed of greater than 220 μm/second were clearly higher in number in the pronase group (n = 35, mean rolling speed 261 μm/second) compared with the trypsin group (n = 19, mean 221 μm/second). Both human (black dots) and porcine (gray dots) BM-MSCs were analyzed. Abbreviations: MSC, mesenchymal stromal/stem cell; ctrl, control; BM, bone marrow.