Fig. 3

Differentiation potential and inflammatory profile of AT-MSCs from responders and non-responders in fistula treatment. AT-MSCs were established from non-responders (n = 15) and responders (n = 12) in AT graft fistula treatment. The cells were examined in undifferentiated and differentiated state in passage 2. Osteoblast differentiation potential of AT-MSCs evaluated by A Alizarin S staining and using quantification of alkaline phosphatase (ALP) activity represented as fold change (F.C.) over non-induced cells (day 7); B and gene expression of RUNX2 and BGALP mRNA levels (n = 12–15); *p < 0.05: non-responders versus responders (two-tailed unpaired Student’s t test). Adipocyte differentiation potential of AT-MSCs evaluated by C Oil red O staining of mature adipocytes (magnification 10x, scale bar 100 μm) and gene expression of PPARG and LPL (n = 12–15); D Gene expression profile of pro-inflammatory (NFKB, IL1B and TNFA) and anti-inflammatory genes (IL10) in non-responder and responder AT-MSCs (n = 12–15). E Gene expression profile of senescence associated secretory phenotype (SASP) (CDKN2A, TPB3, TGFB1, VEGFA, IFNG, IL6) in non-responder and responder AT-MSCs (n = 5); F Gene expression profile of matrix metalloproteinases (MMP2, MMP9) in non-responder and responder AT-MSCs (n = 5). Data are presented as means ± SEM; *p < 0.05, **p < 0.01, ***p < 0.001: non-responders versus responders (two-tailed unpaired Student’s t test)