Journal: Developmental cell

Article Title: Highly efficient methods for generating deletion mutations and F0 embryos that lack gene function in zebrafish

doi: 10.1016/j.devcel.2019.10.004

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: ​ REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Anti-activated Caspase3 BD Biosciences RRID: AB_397274 4D9 (Mouse anti-Engrailed) Monoclonal Antibody Facility, Institute of Neuroscience, University of Oregon RRID: AB_528224 Rabbit polyclonal anti-Prox1 AngioBio RRID: AB_10013720 Goat anti-rabbit lgG-488 Jackson Labs Cat# 111-545-144 Goat anti-rabbit lgG-594 Jackson Labs Cat# 711-585-152 Goat anti-mouse IgG-HRP Jackson Labs Cat# 115-035-003 Chemicals, Peptides, and Recombinant Proteins NEBuffer IM 2.1 NEB Cat# B7202S Duplex buffer IDT Cat# 11-05-01-03 Alt-R® S.p.

Techniques: Recombinant, Amplification, Synthesized, Labeling, Software

Journal: JTCVS Open

Article Title: Therapeutic potential of clinical-grade human induced pluripotent stem cell-derived cardiac tissues

doi: 10.1016/j.xjon.2021.09.038

Figure Lengend Snippet: Engraftment and attenuation of ventricular remodeling after human induced pluripotent stem cell–derived cardiac tissue ( HiCT ) transplantation. A, Representative immunostaining for Ku80 ( brown ) at 4 weeks after surgery. Left and center panels , lower-magnification views. The yellow dotted lines indicate Ku80 + area. (Scale bars: 1000 μm.) Right panel , higher-magnification view. (Scale bar: 100 μm.) B, Quantitative evaluation of the Ku80 + area. The HiCT group showed a significantly larger engraftment area compared with the gelatin hydrogel microsphere–negative [ GHM(-) ] group in both study periods. The upper and lower borders of the box represent the upper and lower quartiles, the middle horizontal line represents the median, and the upper and lower whiskers represent the maximum and minimum values. ∗ P < .05, ∗∗ P < .01, Wilcoxon/Kruskal–Wallis test. Four-week observation: GHM(-), n = 6; HiCT, n = 6; 12-week observation: GHM(-), n = 6; HiCT, n = 6. C and D, Representative immunofluorescence staining at engrafted graft regions. The white dotted line indicates the boundary of the host and graft tissues. C, Cardiac isoform of troponin-T ( cTnT ) ( red ; cardiomyocytes [CMs]), von Willebrand factor ( vWF ) ( green ; endothelial cells [ECs]) and 4',6-diamidino-2-phenylindole ( DAPI ) ( blue ; cell nuclei). (Scale bars: 200 μm [ left ]; 100 μm [ right ]). D, cTnT ( green ; CMs) and DAPI ( blue ; cell nuclei). The white arrow indicates sarcomeric structure. (Scale bars: 100 μm in the left panel; 10 μm in the right panel.) E, Representative Sirius Red staining at 4 weeks after surgery. (Scale bars: 1000 μm.) F, Quantitative evaluations of scar area (%left ventricle [ LV ]). The HiCT group exhibited a significantly smaller scar area compared with the other groups. The upper and lower borders of the box represent the upper and lower quartiles, the middle horizontal line represents the median, and the upper and lower whiskers represent the maximum and minimum values. ∗ P < .05, ∗∗ P < .01 versus sham, † P < .05 versus GHM(-), Wilcoxon/Kruskal–Wallis test, post hoc Steel–Dwass multiple-comparisons test. Four-week observation: sham, n = 11; GHM(-), n = 6; HiCT, n = 6; 12-week observation: sham, n = 8; GHM(-), n = 5; HiCT, n = 6.

Article Snippet: In addition, to evaluate human cells and vascular structure in engraft area, double staining with a rabbit polyclonal anti-Ku80 antibody (Cell Signaling Technology, Danvers, Mass; 1:200) and mouse monoclonal anti-CD31 antibody (Leica Biosystems, Buffalo Grove, Ill; 1:100) was conducted using a conventional ABC method (avidin-biothin-peroxidase complex, ABC-Elite; Vector Laboratories, Burlingame, Calif).

Techniques: Derivative Assay, Transplantation Assay, Immunostaining, Immunofluorescence, Staining

Journal: JTCVS Open

Article Title: Therapeutic potential of clinical-grade human induced pluripotent stem cell-derived cardiac tissues

doi: 10.1016/j.xjon.2021.09.038

Figure Lengend Snippet: Neovascularization after human induced pluripotent stem cell–derived cardiac tissue ( HiCT ) transplantation. A, Representative immunofluorescence staining for von Willebrand factor ( vWF ) ( green ; endothelial cells [ECs]) and 4',6-diamidino-2-phenylindole ( DAPI ) ( blue ; cell nuclei) at the border zone of MI at 4 weeks after surgery. (Scale bars: 100 μm.) B, Quantitative evaluation of vascular density (/mm 2 ) at 4 and 12 weeks after surgery. HiCT transplantation significantly increased vWF + capillaries in close proximity to the graft compared with the sham and gelatin hydrogel microsphere–negative [ GHM(-) ] groups in both study periods. The upper and lower borders of the box represent the upper and lower quartiles, the middle horizontal line represents the median, and the upper and lower whiskers represent the maximum and minimum values. ∗ P < .05 versus sham, † P < .05 versus GHM(-), Wilcoxon/Kruskal–Wallis test, post hoc Steel–Dwass multiple-comparison test. Four-week observation: sham, n = 6; GHM(-), n = 5; HiCT, n = 6; 12-week observation: sham, n = 4; GHM(-), n = 5; HiCT, n = 6. C, Representative light sheet fluorescence microscopy (LSFM) image after 3 days ( left ) and 2 weeks ( right ) of HiCT transplantation. Hoechst 33342 staining ( blue ) indicates transplanted grafts. vWF ( green ) indicates the vascular structure. The white dotted line indicates the boundary of the host and graft tissues. Marked vascular network formation among the grafts was observed at 2 weeks after transplantation. (Scale bars: 700 μm [ left ], 500 μm [ right ]). D, Representative immunohistochemical image at 12 weeks after HiCT transplantation. The black dotted line indicates the boundary of host and graft tissues. Vascular structures among engrafted HiCT include both human (Ku80 + ) and nonhuman, rat cells (Ku80 - , hematoxylin + ). (Scale bars: 100 μm [ left ], 50 μm [ right ]). MI , Myocardial infarction.

Article Snippet: In addition, to evaluate human cells and vascular structure in engraft area, double staining with a rabbit polyclonal anti-Ku80 antibody (Cell Signaling Technology, Danvers, Mass; 1:200) and mouse monoclonal anti-CD31 antibody (Leica Biosystems, Buffalo Grove, Ill; 1:100) was conducted using a conventional ABC method (avidin-biothin-peroxidase complex, ABC-Elite; Vector Laboratories, Burlingame, Calif).

Techniques: Derivative Assay, Transplantation Assay, Immunofluorescence, Staining, Comparison, Fluorescence, Microscopy, Immunohistochemical staining