Abstract 18736: Human Adipose Tissue Inflammation Decreases CD271-Positive Adipose Derived Stem Cells and Deteriorates Therapeutic Angiogenesis of Cell Transplantation for Critical Limb Ischemia
Introduction: Adipose derived stem cells (ADSCs) have been tested in cardiovascular diseases (CVD). However, CVD patients often accompany adipose tissue (AT) inflammation triggered by impaired angiogenesis. Therefore, ADSCs obtained from AT with inflammation might not be suitable for angiogenic cell therapy.
Hypothesis: We hypothesized that AT inflammation would decrease the specific subset of ADSCs and impair the angiogenic efficacy of ADSCs in a hindlimb ischemia model.
Methods and Results: Seventeen patients who underwent cardiac surgery were recruited. Subcutaneous AT was obtained and digested with collagenase to isolate stromal vascular fraction. Flowcytometric analysis was performed to determine cell subsets as follows; 1) CD45+CD14+CD206+ AT macrophages for pro-inflammatory (M1: CD11c+) or alternative (M2: CD11c-) polarization, 2) CD45-CD34+ ADSCs for several subsets previously reported to be therapeutic in other studies (e.g. VEGFR2, CD271). Among studied markers, the frequency of CD271+ /CD45-CD34+ (31.0±3.6%) was negatively correlated with the ratio of M1 to M2 macrophages (0.08±0.02; r=-0.51, p<0.05, n=17). Cultured ADSCs with the lower frequency of CD271+ (0.3-16.8%) showed significantly decreased expression of VEGF compared to those with the higher frequency of CD271+ (46.5-48.6%) in qPCR analysis (relative expressions; 0.03±0.01 vs. 0.25±0.06, p<0.05, n=10). Nude mice were subjected to hindlimb ischemia and one million of human ADSCs at passage 6 were injected into the ischemic muscles 24 hours after resection of left femoral artery. At 2 weeks, neovascularization was evaluated by blood flow recovery (laser Doppler analysis) and capillary density (histological analysis using fluorescent-labeled lectin perfusion). Intramuscular injection of ADSCs with the lower frequency of CD271+ (n=6) in ischemic hindlimb demonstrated impaired neovascularization compared to those with the higher frequency (n=9) in laser Doppler analysis (the ischemic / normal ratio, 28.9±6.3% vs. 55.0±3.5%; p<0.05, n=15) and also in histological analysis (lectin+ endothelial cells, 382±36 vs. 550±81 cells /mm2; p<0.05, n=15).
Conclusions: AT inflammation deteriorates neovascularization induced by ADSCs with decreasing the CD271+ subset.
Author Disclosures: O. Inoue: None. S. Usui: None. S. Takashima: None. T. Hamaoka: None. T. Kitano: None. H. Ootsuji: None. A. Nomura: None. K. Iino: None. T. Kato: None. H. Murai: None. H. Furusho: None. H. Takemura: None. S. Kaneko: None. M. Takamura: None.
- © 2016 by American Heart Association, Inc.