Cd73 pe clone rea804

Post mortem heart and body weights were measured, and hematoxylin/eosin-stained heart, lung, liver, and spleen tissue were studied. Cardiac interstitial fibrosis was evaluated on trichrome-stained sections. We quantified six random hearts from each group, and each heart was sectioned to 10 μm with every 10th section collected. We analyzed 4–13 slices for each group and calculated the percentage of fibrosis with four continuous slices that showed a maximum fibrosis and a coefficient of variation less than 15%. These sections were then scanned with a microscope under 40× magnification. For fibrosis quantification, fibrosis was calculated by count of blue pixels/count of blue and red pixels. The red and blue pixels were distinguished by changing the color threshold with ImageJ software, and the count of these pixels was measured. Color threshold settings for the entire heart (both red and blue pixels) was RGB color space with the triangle thresholding method. The fibrosis (blue pixels) was quantified with pixels with a hue between 150 and 195 by the triangle thresholding method. Heart sections were also stained with an iron staining kit from Sigma-Aldrich to visualize the SIO. The presence of injected hMSCs in the heart was evaluated by immunostaining for CD73-PE (clone REA804), CD90-FITC (clone REA897), and CD105-APC (clone 43A4E1) from Miltenyi Biotech.