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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. 1 — a Schematic illustration of human cardiac tissue fabrication using three cell types, heart extracellular matrix (HEM) hydrogel, and a microfluidic chip. b Hematoxylin and eosin (H&E) staining and DNA content, c Alcian blue staining and glycosaminoglycan (GAG) content, and d Masson’s trichrome (MT) staining and collagen content of native and decellularized porcine heart tissues (scale bars = 50 μm, N = 3, biological replicates, ***p < 0.001). e–g Proteomic analysis of decellularized porcine tissue-derived extracellular matrix (ECM) samples. e ECM composition and the 10 most abundant matrisome proteins in porcine HEM hydrogel samples derived from different batches (A#1, A#2, and A#3) and different donors (A, B, and C) (N = 3, biological replicates). Red letters represent proteins that overlapped between different batches or donors. f Principal component analysis and Pearson’s correlation analysis of total proteins contained in different HEM samples (A#1, A#2, A#3, B, and C). The mean values of three biological replicates were used for analysis. g Percentage of heart tissue-enriched proteins (4-fold higher than in other tissues) among total proteins contained in the porcine HEM A#1 sample (left) and Gene Ontology Biological Processes (GOBP) analysis of the enriched proteins (right). Fisher’s exact test with false discovery rate correction was used for data analysis. h Elastic moduli of HEM hydrogels produced at concentrations of 2, 4, and 6 mg/ml (N = 4, biological replicates, **p < 0.01 and ***p < 0.001). i The projected areas of cardiac tissues fabricated using 2, 4, and 6 mg/ml HEM hydrogel (N = 8, biological replicates). j Immunofluorescent images of cardiomyocyte (CM; cTnT), endothelial cell (EC; CD31), and cardiac fibroblast (CF; VIM) markers in each group of cardiac tissues (scale bars = 200 μm). k Relative mRNA expression levels of CM (MYH7, TNNT2, and NPPA), EC (PECAM1, vWF, and CDH5), and CF (COL1A1 and PDGFRA) markers in each group of cardiac tissues (N = 4, biological replicates, *p < 0.05, **p < 0.01, ***p < 0.001, data are representative of two independent experiments). Cardiac tissues prepared using human induced pluripotent stem cell (hiPSC)-derived CMs, CFs, and human umbilical vein endothelial cells (HUVECs) in (i–k) were cultured for 7 days and used for analysis. Data are presented as means ± S.D. Statistical significance was determined using unpaired two-sided Student’s t-tests in (b–d) and one-way ANOVA with Tukey’s multiple comparisons tests in (h and k). Non-significant statistical differences are indicated as n.s. (p > 0.05).