Correction: Ruchaya et al. Transplantation of Skeletal Muscle-Derived Sca-1+/PW1+/Pax7- Interstitial Cells (PICs) Improves Cardiac Function and Attenuates Remodeling in Mice Subjected to Myocardial Infarction. Cells2022, 11, 4050

Abstract

In the original publication [...].

Figure 1

PIC transplantation improves cardiac function after MI. (A) Schematic diagram of experimental design and echocardiography measurements were taken, before myocardial infarction (MI) (baseline); 7 days, 21 days and 42 days after MI. Post BL echocardiography, animals underwent MI surgery with PBS, MI with PICs or Sham surgery. (B) Changes in cardiac function post-MI. EF = ejection fraction, FS = fractional shortening, SV = stroke volume, LVEED = left ventricular end diastolic dimension, LVESD = left ventricular end systolic dimension. Sham = 8, MI-PICS = 7 and MI-PBS = 7. Data expressed as Mean ± SEM, * p < 0.05 vs. Sham, # p < 0.05 vs. MI-PBS.

Figure 2

PIC transplantation attenuates cardiac remodelling after MI. (A) Fibrosis detection using hematoxylin van geisen (HVG) staining in Sham = 8, MI-PICS = 7 and MI-PBS = 7, pink/red staining indicates fibrosis. Data expressed as Mean ± SEM of the fibrotic area over the total area of the left ventricle (LV), * p < 0.05 vs. Sham, # p < 0.05 vs. MI-PICs, scale bar = 500 µM. (B) Representative micrograph depicting the risk region (RR) and non-risk region (non-RR) in the LV. Scale bar 100 µM. (C) % CC3+ apoptotic cardiomyocytes in the RR of the LV, representative image of Sham, MI-PBS and MI-PICs. Green; wheat germ agglutinin (WGA); red; cleave caspase 3 (CC3); blue; DAPI. Arrowheads show CC3+ cardiomyocytes in MI-PBS. Arrowheads show non-cardiomyocyte CC3+ cells in MI-PICs. Sham = 8, MI-PICS = 7 and MI-PBS = 6. Scale bar = 25 µM. Data are Mean ± SEM. * p < 0.05 vs. Sham, # p < 0.05 vs. MI-PICs. (D) Representative confocal micrographs of the RR in Sham = 8, MI-PICs = 6 and MI-PBS = 7 groups, WGA (green) and DAPI (blue). Scale bar = 500 µM. Graphs showing the cross-sectional area and circumference of cardiomyocytes in the RR of the LV. Data expressed as Mean ± SEM, * p < 0.05 vs. Sham, # p < 0.05 vs. MI-PICs.

Figure 3

PICs engraft in the infarcted myocardium. (A) Percent engraftment of GFP+ PICs 6 weeks post-transplantation, represented as a percentage of the total number of DAPI cells in the risk region (RR) and non-risk region (non-RR). Data are Mean ± SEM in 20 FOV per mouse. (B–D) Representative confocal micrographs of GFP+ PICs (green) expressing markers of cardiomyocyte ((B), Red; α-sarcomeric actin), smooth muscle ((C), Red; calponin) and endothelial ((D), Red, vWF) cells. DAPI stain nuclei blue. Scale bar = 50 µM. (E) Quantification of the GFP+ PICs expressing markers of the cardiac tr-lineage in MI-PICs group n = 6. Data are Mean ± SEM.

Figure 4

PICs transplantation increases proliferation and number of BrdU⁺ cardiomyocytes. (A) Representative confocal micrographs in the RR of the LV of BrdU (Red) positive cells. DAPI stain nuclei blue. Scale bar = 10 µM. Arrows indicate positive staining colocalisation. (B) Quantification of the number of proliferating BrdU-positive cells in the RR in Sham = 8, MI-PBS = 7 and MI-PICs = 6 groups. Data are Mean ± SEM of the total number of BrdU⁺ cells over the total number of cells (DAPI), * p < 0.05 vs. sham, # p < 0.05 vs. MI-PBS. (C–E) Representative confocal micrographs in the RR of the LV of BrdU (Green) positive, α-sarcomeric actin-expressing ((C), Red), calponin-expressing ((D), Red) or vWF-expressing ((E), Red) cells. DAPI stain nuclei blue. Scale bar = 10 µM; arrows indicate positive staining colocalisation. (F) Quantification of the number of BrdU⁺ trilineage cardiac expressing cells. Data are Mean ± SEM. (G) Representative image of BrdU⁺ cardiomyocyte (WGA) in MI-PICs. DAPI stain nuclei blue. Arrows indicate cells that show colocalisation. Sham = 5, MI-PICS = 5 and MI-PBS = 5. Scale bar = 10 µM. (H) Percent number of BrdU⁺ cardiomyocytes in the RR. Data are Mean ± SEM. * p < 0.05 vs. sham, # p < 0.05 vs. MI-PBS.

Figure 5

PICs have a pro-reparative secretome. (A) Pro-survival and regenerative transcript expression of PICs. Data are relative expression to housekeeping genes GAPDH, ß-actin and B2M. Gene profile relative expression levels are depicted as, >500, 500–10 and <10. Data are Mean ± SD, n = 3. The arrows indicate the genes that were further investigated at the protein level. (B) Protein quantification using ELISA; concentration of mouse IGF-1, VEGF, HGF, Follistatin and TGFß1 (pg/mL) in mPIC conditioned media (CM) relative to unconditioned (control) media. Data are Mean ± SD, n = 3. (C) Matrigel tube formation assay. HUVECs were cultured for 15 h in 96-well plates coated with matrigel. Data are presented as the Mean ± SEM. Positive control, HUVEC media, Negative Control, DMEM and Conditioned media; is serum free DMEM conditioned with PICs media * p < 0.05 vs. Positive control, # p < 0.05 vs. Negative control.