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. 2022 Jan 20;8:806215. doi: 10.3389/fcvm.2021.806215

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Copyright © 2022 Kawai, Tohyama, Arai, Tamura, Soma, Fukuda, Shimizu, Nakayama and Kobayashi.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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The tubular engineered heart tissues (T-EHTs) after 4 w of transplantation. (A,B) The T-EHT in vivo, highlighted in fluorescent imaging (black arrows). (C,D) The T-EHT is being explanted with surrounding tissues, and its fluorescence imaging record (black arrows). (E) Beating analysis of explanted T-EHT. The T-EHT showed spontaneous beating. (F) Beating analysis of explanted T-EHT with electrical stimulation. The T-EHT was stimulated with bipolar electrical pulses at 15 V and 1 Hz or 2 Hz for 10 ms. Under electrical stimulation, the beating rate increased to 1.4 Hz at 1 Hz stimulation and 1.8 Hz at 2 Hz stimulation and reduced to 1.1 Hz and 1.6 Hz after stimulation, respectively. Y-axis represents the amounts of movement (in pixels) of the T-EHT recorded using a digital camera. (G–L) Histological staining of T-EHT and its high magnification image. (G) Hematoxylin and eosin (HE) staining. (H) Myosin light chain 2v (MLC2v) and N-cadherin. (I) CD31 and cardiac troponin T (cTnT). (J) α-actinin and vimentin. (K) MLC2v and cTnT. (L) MLC2v and myosin light chain 2a (MLC2a). The HE staining shows engraftment of T-EHT (black arrowheads) and vascularization (black arrow) (G). White arrowheads in the immune staining show T-EHT. Striation of cardiomyocytes are observed in MLC2v, cTnT, and α-actinin staining (H–K). Compared to the T-EHT observed 1 w after transplantation, the number of MLC2a-positive cells is reduced 4 w after transplantation (L).