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. 2024 Mar 22;15:2564. doi: 10.1038/s41467-024-46928-y

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© The Author(s) 2024

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 2 — a Schematic illustrations of the fabrication of cardiac tissues using HEM hydrogel, conventional commercial hydrogels, or U-bottom plates. b Bright-field images and c projected areas of human cardiac tissues fabricated using U-bottom plates (No ECM [extracellular matrix]), collagen, fibrin, growth factor reduced (GFR)-Matrigel, embryonic stem cell (ESC)-qualified Matrigel, and HEM hydrogel (scale bar = 0.5 mm, N = 9, biological replicates, ***p < 0.001 versus HEM group). d Representative immunofluorescent images and e quantification of the areas positive for cardiomyocyte (CM; cTnT), endothelial cell (EC; CD31), and cardiac fibroblast (CF; VIM) markers in each group of cardiac tissues (scale bars = 0.5 mm, N = 4, biological replicates, *p < 0.05 and ***p < 0.001 versus HEM). The cTnT-positive areas were quantified using immunofluorescent images in (d). f Quantitative PCR (qPCR) analysis of relative mRNA expression levels of CM (MYH7), EC (PECAM1), and CF (COL1A1) markers in each group of cardiac tissues (N = 4, biological replicates, **p < 0.01 and ***p < 0.001 versus HEM group). g Schematic illustrations of fabrication of cardiac tissues using HEM hydrogel and other tissue-derived ECM hydrogels. h Bright-field images and i projected areas of cardiac tissues fabricated using decellularized intestine, stomach, and muscle-derived ECM hydrogels and HEM hydrogel (scale bar = 0.5 mm, N = 10, biological replicates, ***p < 0.001 versus HEM group). j Representative immunofluorescent images and k quantification of the areas positive for CM (cTnT), EC (CD31), and CF (VIM) markers in each group of cardiac tissues (scale bars = 0.5 mm, N = 5, biological replicates, **p < 0.01 versus HEM group). The cTnT-positive areas were quantified using immunofluorescent images in (j). l qPCR analysis of relative mRNA expression levels of CM (MYH7 and TNNT2), EC (PECAM1 and vWF), and CF (COL1A1 and PDGFRA) markers in each group of cardiac tissues (N = 4, biological replicates, **p < 0.01 and ***p < 0.001 versus HEM group). Cardiac tissues prepared with human induced pluripotent stem cell (hiPSC)-derived CMs, CFs, and human umbilical vein endothelial cells (HUVECs) were cultured for 7 days and used for analysis. Data are presented as means ± S.D. Statistical significance was determined using one-way ANOVA with Tukey’s multiple comparisons tests in (c, e, f, i, k, and l).