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Transport viable heart tissue at physiological temperature yielded higher human cardiomyocytes compared to the conventional temperature

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Abstract

This study investigated the optimum transport condition for heart tissue to recover single-cell cardiomyocytes for future in-vitro or in-vivo studies. The heart tissues were obtained from removing excessive myocardium discharged during the repair surgery of an excessive right atrial hypertrophy due to a congenital disease. The transportation temperature studied was the most used temperature (4 °C) or the conventional condition, compared to a physiological temperature(37 °C). The heart tissues were transported from the operating theatre to the lab maintained less than 30 min consistently. Single-cell isolation was enzymatically and mechanically performed using collagenase-V (160 U/mg) and proteinase-XXIV (7-14 U/mg) following the previously described protocol. The impact of temperature differences was observed by the density of cells harvested per mg tissue, cell viability, and the senescence signals, identified by the p21, p53 and caspase-9 mRNA expressions. Results the heart tissue transported at 37 °C yielded significantly higher viable cell density (p < 0.01) yielded viable cells significantly higher density (p < 0.01) than the 4 °C; 2,335 ± 849 cells per mg tissue, and 732 ± 425 cells per mg tissue, respectively. The percentage of viable cells in both groups showed no difference. Although the 37 °C group expressed the apoptosis genes such as p21, p53 and caspase9 by 2.5-, 5.41-, 5-fold respectively (p > 0.05). Nonetheless, the Nk×2.5 and MHC genes were expressed 1,7- and 3.56-fold higher than the 4 °C. and the c-Kit+ expression was 17.56-fold, however, statistically insignificant. Conclusion When needed for single-cell isolation, a heart tissue transported at 37 °C yielded higher cell density per mg tissue compared to at 4 °C, while other indicators of gene expressions for apoptosis, cardiac structural proteins, cardiac progenitor cells showed no difference. Further investigations of the isolated cells at different temperature conditions towards their proliferation and differentiation capacities in a 3-D scaffold would be essential.

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Data availability

The datasets generated during and/or analysed during the current study are not publicly available due to the confidentiality of patients’ data but are available from the corresponding author on reasonable request.

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Funding

This study was supported by Grant No. NKB-581/UN2.RST/HKP.05.00/2020 from the University of Indonesia.

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Authors and Affiliations

  1. Department of Cardiovascular and Thoracic Surgery, Faculty of Medicine, Universitas Indonesia, 10430, Jakarta, Indonesia

    Muhammad Arza Putra,  Ardiansyah, Chaidar Muttaqin, William Makdinata & Idrus Alwi

  2. Human Reproduction Infertility and Family Planning Research Center, Indonesia Medical Education and Research Institute, 10430, Jakarta, Indonesia

    Normalina Sandora, Raisa Nauli, Tyas Rahmah Kusuma & Nur Amalina Fitria

  3. Infectious Disease and Immunology Research Center, Indonesia Medical Education and Research Institute, 10430, Jakarta, Indonesia

    Suwarti

  4. Stem Cell and Tissue Engineering Research Center, Indonesia Medical Education and Research Institute, 10430, Jakarta, Indonesia

    Retno Wahyu Nurhayati

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  1. Muhammad Arza Putra
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Correspondence to Normalina Sandora.

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All methods in this study have been reviewed and approved by the Ethical Committee of Universitas Indonesia with Ethical Approval no. KET.483/UN2.F1/ETIK/PPM.00.02/2019.

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Putra, M.A., Sandora, N., Suwarti et al. Transport viable heart tissue at physiological temperature yielded higher human cardiomyocytes compared to the conventional temperature. Cell Tissue Bank 23, 717–727 (2022). https://doi.org/10.1007/s10561-021-09978-w

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