Cardioprotective Potential of Human Endothelial-Colony Forming Cells from Diabetic and Nondiabetic Donors

• Published in: Cells (Basel, Switzerland) Switzerland), 2020-03, Vol.9 (3), p.588
• Main Authors: Deutsch, Marcus-André, Brunner, Stefan, Grabmaier, Ulrich, David, Robert, Ott, Ilka, Huber, Bruno C
• Format: Article
• Language: English
• Subjects: adult stem cells; angiogenesis; Animals; cardiac regeneration; Cardiotonic Agents - metabolism; cardiovascular diseases; Diabetes Mellitus - physiopathology; Disease Models, Animal; Endothelial Cells - metabolism; Humans; Male; Mice; Mice, SCID; Myocardial Infarction - physiopathology; myocardial infraction; Tissue Donors
• ISSN: 2073-4409; 2073-4409
• DOI: 10.3390/cells9030588

The potential therapeutic role of endothelial progenitor cells (EPCs) in ischemic heart disease for myocardial repair and regeneration is subject to intense investigation. The aim of the study was to investigate the proregenerative potential of human endothelial colony-forming cells (huECFCs), a very homogenous and highly proliferative endothelial progenitor cell subpopulation, in a myocardial infarction (MI) model of severe combined immunodeficiency (SCID) mice. CD34+ peripheral blood mononuclear cells were isolated from patient blood samples using immunomagnetic beads. For generating ECFCs, CD34 cells were plated on fibronectin-coated dishes and were expanded by culture in endothelial-specific cell medium. Either huECFCs (5 × 10 ) or control medium were injected into the peri-infarct region after surgical MI induction in SCID/beige mice. Hemodynamic function was assessed invasively by conductance micromanometry 30 days post-MI. Hearts of sacrificed animals were analyzed by immunohistochemistry to assess cell fate, infarct size, and neovascularization (huECFCs = 15 vs. control = 10). Flow-cytometric analysis of enzymatically digested whole heart tissue was used to analyze different subsets of migrated CD34+ /CD45+ peripheral mononuclear cells as well as CD34 /CD45 cardiac-resident stem cells two days post-MI (huECFCs = 10 vs. control = 6). Transplantation of human ECFCs after MI improved left ventricular (LV) function at day 30 post-MI (LVEF: 30.43 ± 1.20% vs. 22.61 ± 1.73%, < 0.001; ΔP/ΔT 5202.28 ± 316.68 mmHg/s vs. 3896.24 ± 534.95 mmHg/s, < 0.05) when compared to controls. In addition, a significantly reduced infarct size (50.3 ± 4.5% vs. 66.1 ± 4.3%, < 0.05) was seen in huECFC treated animals compared to controls. Immunohistochemistry failed to show integration and survival of transplanted cells. However, anti-CD31 immunohistochemistry demonstrated an increased vascular density within the infarct border zone (8.6 ± 0.4 CD31 capillaries per HPF vs. 6.2 ± 0.5 CD31 capillaries per HPF, < 0.001). Flow cytometry at day two post-MI showed a trend towards increased myocardial homing of CD45 /CD34 mononuclear cells (1.1 ± 0.3% vs. 0.7 ± 0.1%, = 0.2). Interestingly, we detected a significant increase in the population of CD34 /CD45 /Sca1 cardiac resident stem cells (11.7 ± 1.7% vs. 4.7 ± 1.7%, < 0.01). In a subgroup analysis no significant differences were seen in the cardioprotective effects of huECFCs derived from diabetic or nondiabetic patients. In a murin