Higher Improvement of Cardiac Function Following Myocardial Infarction using Menstrual Blood Stromal/Stem Cells (MenSCs) Suspended in Conditioned Medium versus Conditioned Medium Alone in Rat Model

Figures

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  Figure 1.. Evaluation of MenSCs by flow cytometry and cardiac parameters by echocardiography at days 7 and 28 post-MI. A) CD markers are demonstrated (gray and white curves). B) % Fractional shortening (FS), C) % Ejection fraction (EF), * p<0.05; ** p<0.001.

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  Figure 2.. Histopathological evaluation of cardiac function at days 7 & 28 post MenSCs administration; A & B) Section from sham group shows normal architecture at day 7, C) In MI group, the ischemic region is mainly occupied by granulation tissue (arrow) at d ay 7, D) Higher magnification of a slide belonged to MI group shows a greater cellular granulation tissue (rectangle) and cardiomyonecrosis (arrow), E) In CM group, the main structure is granulation tissue (circle) at day 7, F) Higher magnification of pervious slide, notes to high cellularity within granulation tissue (rectangle), and necrotic cardiomyocytes (arrow), G) In MenSCs+ CM group, many preserved cardiomyocytes (circle) could be seen in the ischemic area at day 7, H) Higher magnification of pervious slide, note to inflammatory cells infiltration (ar row) among preserved muscle fibers (rectangle), I & J) Normal histology of section belonged to sham group at day 28, K) Note to myocardial scar tissue bulged (arrow) at day 28, L) Higher magnification of pervious image, notes to cartilaginous metaplasia (star) in the center o f scar tissue, M) Note to mature granulation tissue (green arrow) and cartilaginous metaplasia (black…

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  Figure 3.. A) Evaluation of myocardial infarction size at day 28 post-MI, representative transversal sections of heart and Masson's trichrome staining for infarct size measurement. Scar tissue is colored in blue and healthy myocardium in red. Scale bar=1.5 mm, B) Percentage of circumferential infarct size (MI size), *p<0.05.

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  Figure 4.. IHC staining of α-sarcomeric actin, connexin-43 at days 7 & 28 and tracking of transplanted MenSCs into the infarcted zone by the anti-human mitochondrial antibody. A & B) Note to noticeable expression of α-sarcomeric actin in cardiomyocytes belonged to sham group (star), C & D) No expression of α-sarcomeric actin is seen in scar tissue of MI group, E & F) No expression is detected in infarct zone of CM group, G & H) Significant expression of α-sarcomeric actin is detected in survived cardiomyocytes in MenSCs + CM group (star), I & J) Sham group, connexin-43 expression at the intercalated disks is evident (rectangle), K & L) MI group; there is no expression in scar tissue, M & N) In CM group, no expression is detectable, O & P) Notice connexin-43 expression at the intercalated disks in preserved cardiomyocytes in MenSCs+CM group (rectangle), Q) 28 days after trans-epicardial injection of MenSCs, the transfer of human mitochondria is successfully detectable in cardiomyocytes in border zone (arrow), (IHC, Scale bar = A-Q: 50 μm).

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  Figure 5.. IHC staining of type I and II collagen in the myocardium at day 28 (A & B) No expression of Col I, (C & D) In MI group, no expression of Col I is detected in cartilaginous metaplastic site, (E) Note to limited expression of this marker by some chondrocyt es in metaplastic site in CM group (arrow), (F) Higher magnification of pervious slide (arrow), (G & H) The expression of Col I in MenSCs+CM gr oup occurs in scar tissue, (I & J) Sham group, no expression of Col II, (K) Note to expression of Col II by chondr ocytes in cartilaginous metaplastic site in cases with no-treated MI (rectangle), (L) Higher magnification of pervious slide (arrow), (M) The positive expression of Col II was detecta ble in metaplastic region of the case that received only CM (rectangle), (N) Higher magnification of pervious slide (arrow), (O & P) There is no detectable expression of Col II in scar tissue of MenSCs+CM group (IHC, Scale bar = A, C, E, G, I, K, M and O: 200 μm; B, D, F, H, J, L, N and P: 50 μm). *p<0.05, p<0.01, and#p<0.001.

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  Figure 6.. IHC staining of NF-κB at day 28. (A & B) No expression of NF-κB is seen in sham group, (C) Note to significant ex-pression of this marker in cardiomyocytes of border zone (circle), and cartilaginous tissue (rectangle) of scar zone in MI group (rectangle), (D) Higher magnification of pervious slide, note to expression of this marker by chondrocytes (rectangle), (E & F) Noticeable expression of N F-κB in metaplastic region (rectangle) in group that received CM, (G & H) Slight expression of NF-κB by survived cardiomyocytes in MenSCs+CM-received group (re-ctangle) (IHC, Scale bar=A, C, E, G: 200 μm; B, D, F, H: 50 μm).

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  Figure 7.. The role of MenSCs+CM in cardiac function.