Figure 1
The aging-associated ROS and impaired functions of elderly donor-derived AT-MSCs to decrease the necrotic area of the flap mouse model. (A) The ROS expression in infant and elderly AT-MSCs. (n = 5). (B) The mRNA expression of pro-inflammatory factors in infant and elderly AT-MSCs (n = 5). (C) The mRNA expression of wound healing-related growth factors in infant and elderly AT-MSCs (n = 5). (D) The migration scratch assay of infant and elderly AT-MSCs, magnification 4 ×. Bar indicates 200 μM (n = 5). (E) The transwell assay of EPCs and ECs co-cultured with infant and elderly AT-MSCs magnification 10 ×. Bar indicates 200 μM (n = 5). (F) The tube formation of EPCs and ECs in the infant and elderly AT-MSC-conditioned medium, magnification 4 ×. Bar indicates 200 μM (n = 5). (G) Transplantation of infant and elderly AT-MSCs to an in vivo streptozotocin-induced diabetic ischemic flap mouse model (n = 5). (H) Immunohistochemical staining with PE-labeled anti-CD45 and Mac 1 on the third day and CD-31 on the seventh day of transplantation of the necrotic areas. The brown dots indicate positive cells, magnification 10 ×. Bar indicates 200 μM (n = 5). (I) The expression of ROS in elderly AT-MSCs in the presence of antioxidants (n = 5). (J) The cellular senescence of elderly AT-MSCs in the presence of antioxidants (n = 5). (K) Transplantation of elderly AT-MSCs and antioxidant-treated elderly AT-MSCs to an in vivo diabetic ischemic flap mouse model. Edaravone (Eda, 20 µM) and N-acetylcystein (NAC 2 mM) were used as antioxidants with a treatment time of 24 h (n = 5). In all above experiments, infant AT-MSCs and elderly AT-MSCs were derived from 5 different donors, respectively (n = 5). AT-MSCs were at passage 3 to passage 8, and the comparison was conducted with infant AT-MSCs and elderly AT-MSCs at the same passage number. The data represent the mean ± SD, ***P < 0.001, **P < 0.01, *P < 0.05, ns no significance. The experiments were performed in triplicate.
