Fig. 1
Coagulant activity of xeno- and serum-cultured MSCs and their expression of pro- and anticoagulant factors. a MSCs derived from AT, BM, DP, and UC as well as HUVECs and PBMNCs (n = 3 each) were prepared in NaCl and RL and supplemented with plasma from healthy donors (n = 6) and CaCl2 to measure the time required to form fibrin clots. MSCs exhibited the highest coagulant activity, followed by HUVECs, PBMNCs and the negative control without cells. b Gene expression analysis of TF revealed the highest expression in UC-MSCs, moderate levels in AT- and DP-MSCs, and low expression in BM-MSCs and HUVECs (UC- and DP-MSCs: n = 10 and other cell types: n = 3). c, d TF protein expression was analyzed by flow cytometry. A representative sample of UC-MSCs demonstrated coexpression of TF/CD142 and MSC-positive markers, including CD90, CD73, and CD105 (c). The frequency of TF+ cells and CD142 MFI were analyzed, showing a high level of this factor in UC- and AT-MSCs, lower levels in DP- and BM-MSCs, and negative expression in HUVECs (d). e–g Gene expression was analyzed for the procoagulant factor COL1A1 and the anticoagulant factors TFPI and PTGIR. UC-MSCs expressed significantly higher COL1A1 than AT- and BM-MSCs and HUVECs (e). In terms of the anticoagulant factors TFPI (f) and PTGIR (g), UC- and AT-MSCs and HUVECs displayed higher expression than DP- and BM-MSCs. h The exposure of the negatively charged phosphatidylserine (PS) in the outer membrane of UC- and DP-MSCs was analyzed using an anti-Annexin V antibody. A subset of UC- and DP-MSCs in culture (continuously cultured cells) were positive for Annexin V