Therapeutic effects of adipose derived fresh stromal vascular fraction-containing stem cells versus cultured adipose derived mesenchymal stem cells on rescuing heart function in rat after acute myocardial infarction

Abstract

We tested the hypothesis that adipose-derived fresh stromal vascular fraction (SVF) is non-inferior to conventional adipose-derived mesenchymal stem cell (ADMSC) therapy for improving left-ventricular ejection fraction (LVEF) in rat after acute myocardial infarction (AMI). Male-adult SD rats (n = 48) were categorized into group 1 (sham control), AMI, AMI + ADMSCs (1.2 × 106) cells] and AMI + SVF (1.2 × 106) cells]. Flow cytometric and qPCR analyses showed that the expressions of surface biomarkers for endothelial progenitor cells, and cardiac-stem cells were significantly higher in the SVF population than in the ADMSC population, whereas MSCs showed a reversed pattern between these two groups (all P < 0.001). By day-42 after AMI, LVEF was highest in SC, lowest in AMI, and significantly higher in AMI + SVF than in AMI + ADMSCs (P < 0.0001). Protein expression indicating angiogenesis, anti-inflammatory/anti-apoptotic, mitochondrial/bioenergy-integrity and antifibrotic biomarkers showed an identical pattern, whereas protein expressions for inflammatory, apoptotic and pressure-overload/heart failure biomarkers exhibited an opposite pattern to LVEF among the four groups (all P < 0.001). Histopathology displayed that LV infarction/fibrotic area/collagen-deposition areas, cellular expressions of DNA-damage, and inflammatory biomarkers exhibited an opposite pattern, whereas cellular expressions of endothelial/gap-junction biomarkers showed an identical pattern to LVEF among the four groups (all P < 0.0001). Cellular expression of angiogenesis biomarkers significantly and progressively increased from groups 1 to 4 (all P < 0.0001). In conclusion, SVF may be better than ADMSC at improving LVEF in rat after AMI.

Keywords: Acute myocardial infarction; adipose-derived mesenchymal stem cells; fresh stromal-vascular fraction containing stem cells; heart function.

Conflict of interest statement

None.

Figures

Figure 1

Gene expressions of EPCs, ECs, MSC and cardiac stem cells in SVF and ADMSC and illustrating the higher subpopulation of stem cells in SVF, and small vessel density and muscularization in LV infarct area by day 42 after AMI induction. Upper panel: A. mRNA expressions of CD31, * vs. †, P

Figure 9

Cellular expressions of DNA-damaged and gap-junction biomarkers in LV myocardium by day 42 after AMI induction. A-D. Illustrating the immunofluorescent (IF) microscopic finding (400 ×) for identification of γ-H2AX+ cells (pink color). Scale bars in right lower corner represent 20 µm. E. Analytical result of percentage of γ-H2AX+ cells, * vs. other group with different symbols (†, ‡, §), P

Figure 2

The microscopic findings of infarct area, fibrotic area, condensed collagen deposition area and cellular expression of heart failure biomarker in left ventricular myocardium by day 42 after AMI induction. Upper panel: A-D. Illustrating H.E. stain (100 ×) for identification of infarct area (pink-white color). a-d. Illustrating the cross section of left ventricle for identification of infarct area. E. Analytical results of the infarct area, * vs. other group with different symbols (†, ‡, §), P

Figure 3

Protein expressions of angiogenesis, apoptotic, anti-apoptotic, fibrotic and anti-fibrotic biomarkers in left ventricular myocardium by day 42 after AMI induction. A. Protein expression of endothelial nitric oxide synthase (eNOS), * vs. other group with different symbols (†, ‡, §), P

Figure 4

Protein expressions of inflammatory, mitochondrial-damaged, mitochondrial/energy-integrity and cardiac hypertrophic/heart failure biomarkers in LV myocardium by day 42 after AMI induction. A. Protein expressions of matrix metalloproteinase (MMP)-9, * vs. other group with different symbols (†, ‡, §), P

Figure 5

Cardiac stem cells in LV myocardium by day 42 after AMI induction. A-D. Illustrating immunohistochemical (IHC) microscopic finding (400 ×) for identification of Sca-1+ cells (gray color). E. Analytical results of number of Sca-1+ cells, * vs. other group with different symbols (†, ‡, §), P

Figure 6

Inflammatory biomarkers in LV myocardium by day 42 after AMI induction. (A-D) Illustrating immunofluorescent (IF) microscopic finding (400 ×) for identification of CD14+ cells (green color). Red color in (C and D) indicated implanted ADMSC and SVF in myocardium, respectively. (E) Analytical result of percentage of CD14+ cells, * vs. other group with different symbols (†, ‡, §), P

Figure 7

Cellular expression of endothelial cell biomarkers in LV myocardium by day 42 after AMI induction. (A-D) Illustrating the immunofluorescent (IF) microscopic finding (400 ×) for identification of CD31+/vov Willebran Factor (vWF)+ cells (i.e., double stain; CD31-green color; vWF-red color). (E) Analytical result of percentage of CD31+/vWF+ cells, * vs. other group with different symbols (†, ‡, §), P

Figure 8

Cellular expressions of endothelial progenitor cell (EPC) biomarkers in LV myocardium by day 42 after AMI induction. (A-D) Illustrating the immunofluorescent (IF) microscopic finding (400 ×) for identification of stromal-cell derived factor (SDF)-1α+ cells (green color). Red color in (C and D) indicated implanted ADMSC and SVF in myocardium, respectively. (E) Analytical result of percentage of SDF-1α+ cells, * vs. other group with different symbols (†, ‡, §), P