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Characterisation of the single-cell human cardiomyocytes taken from the excess heart tissue of the right ventricular outlet in congenital heart disease

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Abstract

Cardiovascular disease is the second highest cause of death across the globe. Myocardial infarction is one of the heart diseases that cause permanent impairment of the heart wall leads to heart failure. Cellular therapy might give hope to regenerate the damaged myocardium. Single cells isolated from an excess heart tissue obtained from the correction of the right ventricular hypertrophy in patients with Tetralogy of Fallot for future heart study were investigated. Methods: Once resected, the heart tissues were transported at 37 °C, in Dulbecco's Modified Eagle's medium/ DMEM (4.5 g.L−1, antibiotic–antimycotic 3x, PRP10% (v/v)), to reach the lab within 30 min, weighted and grouped into less than 500 mg and more than 1000 mg (n = 4). Each sample was digested with 250 U.mL−1 Collagenase type V and 4U.mL−1 Proteinase XXIV in the MACS™ C-tube (Milltenyi, Germany), then dissociated using the MACS™ Octo Dissociator with Heater (Milltenyi, Germany) for 60 min at 37 °C. Results: All cells isolated were rod-shaped cells; viability was up to 90%. The cell density obtained from the 500 mg group were 4,867 ± 899 cells.mg−1 tissue weight, significantly higher compared to the 1,000 mg group; had 557 ± 490 cells.mg−1 tissue weight (mean of (n = 3) ± 95% C.l). The isolated cells were analyzed using FACs BD Flowcytometer, expressed cTnT + 13.38%, PECAM-1 + /VCAM-1- 32.25%, cKit + 7.85%, ICAM + 85.53%, indicating the cardiomyocyte progenitor cells. Conclusion: Cardiomyocytes taken from the wasted heart tissue might be a candidate of cardiomyocytes source to study interventions to the heart as it contained up to 13.38% cardiomyocytes, and 32.25% of cardiac progenitor cells. Moreover, perhaps when cardiac cell therapy needs autologous cardiomyocytes, less than 500 mg tissue weight can be considered as sufficient.

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Data availability

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

References

  • Au-Bhattacharya S, Au-Burridge PW, Au-Kropp EM, Au-Chuppa SL, Au-Kwok W-M, Au-Wu JC, Au-Boheler KR, Au-Gundry RL (2014) High Efficiency Differentiation of Human Pluripotent Stem Cells to Cardiomyocytes and Characterization by Flow Cytometry. JoVE, e52010

  • Brand S, Gerber M, Beck J, Hatzinger M, Pühse U, Holsboer-Trachsler E (2010) High exercise levels are related to favorable sleep patterns and psychological functioning in adolescents: a comparison of athletes and controls. J Adolescent Health 46:133–141

  • Bulluck H, Fröhlich GM, Nicholas JM, Mohdnazri S, Gamma R, Davies J, Sirker A, Mathur A, Blackman D, Garg P, Moon JC, Greenwood JP, Hausenloy DJ (2019) Mineralocorticoid receptor antagonist pre-treatment and early post-treatment to minimize reperfusion injury after ST-elevation myocardial infarction: the MINIMIZE STEMI trial. Am Heart J 211:60–67

    Article  CAS  Google Scholar 

  • Butts RJ, Scheurer MA, Atz AM, Moussa O, Burnette AL, Hulsey TC, Savage AJ (2014) Association of human leukocyte antigen donor–recipient matching and pediatric heart transplant graft survival. Circulation Heart Failure 7:605–611

    Article  CAS  Google Scholar 

  • Cheung JY, Leaf A, Bonventre J (1984) Determination of isolated myocyte viability: staining methods and functional criteria. Springer, Adult heart muscle cells

    Google Scholar 

  • Entman ML, Smith CW (1994) Postreperfusion inflammation: a model for reaction to injury in cardiovascular disease. Cardiovasc Res 28:1301–1311

    Article  CAS  Google Scholar 

  • FAVREAU-LESSARD, A. J. & SAWYER, D. B. 2018. Cardiac Regeneration and Stem Cells as Therapy for Heart Disease. In: VASAN, R. S. & SAWYER, D. B. (eds.) Encyclopedia of Cardiovascular Research and Medicine. Oxford: Elsevier

  • Frangogiannis NG, Smith CW, Entman ML (2002) The inflammatory response in myocardial infarction. Cardiovasc Res 53:31–47

    Article  CAS  Google Scholar 

  • Ghani L, Susilawati MD, Novriani H (2016) Faktor risiko dominan penyakit jantung koroner di Indonesia. Buletin Penelitian Kesehatan 44:153–164

    Google Scholar 

  • Graesser D, Solowiej A, Bruckner M, Osterweil E, Juedes A, Davis S, Ruddle NH, Engelhardt B, Madri JA (2002) Altered vascular permeability and early onset of experimental autoimmune encephalomyelitis in PECAM-1–deficient mice. J Clin Investig 109:383–392

    Article  CAS  Google Scholar 

  • Grunow B, Wenzel J, Terlau H, Langner S, Gebert M, Kruse C (2011) In vitro developed spontaneously contracting cardiomyocytes from rainbow trout as a model system for human heart research. Cell Physiol Biochem 27:1–12

  • HORDIJK, P. L. (2006) Endothelial signalling events during leukocyte transmigration. FEBS J 273:4408–4415

    Article  Google Scholar 

  • IRAWAN, C., SANTOSO, T., ATMAKUSUMAH, D., ALWI, I., KOSASIH, A., AZIZ, A., ABDULMUTHALIB, A., LYANA, L., INGGRIANI, S. & SAPUTRA, A. 2016. Hemostatic Status of Pre and Post Intracoronary Injection of Peripheral Blood Stem Cells in Patients with Recent Myocardial Infarction. Acta Medica Indonesiana, 46

  • JARCHO, J., NAFTEL, D., SHROYER, T., KIRKLIN, J., BOURGE, R., BARR, M., PITTS, D. & STARLING, R. 1994. Influence of HLA mismatch on rejection after heart transplantation: a multiinstitutional study. The cardiac transplant research database group. J Heart Lung Transplant Offic Pub Int Soc Heart Transplant, 13, 583–95; discussion 595–6

  • Jopling C, Suñé G, Faucherre A, Fabregat C, IZPISUA BELMONTE, J. C. (2012) Hypoxia induces myocardial regeneration in zebrafish. Circulation 126:3017–3027

    Article  Google Scholar 

  • Khush KK, Menza R, Nguyen J, Zaroff JG, Goldstein BA (2013) Donor predictors of allograft use and recipient outcomes after heart transplantation. Circulat Heart Failure 6:300–309

    Article  Google Scholar 

  • Biro Komunikasi Dan Pelayanan Masyarakat, K. KR (2017) Penyakit Jantung Penyebab Kematian Tertinggi, Kemenkes Ingatkan CERDIK. 29 Juli 2017 ed. Jakarta: sKementerian Kesehatan RI

  • Kwee L, Baldwin HS, Shen HM, Stewart CL, Buck C, Buck CA, Labow MA (1995) Defective development of the embryonic and extraembryonic circulatory systems in vascular cell adhesion molecule (VCAM-1) deficient mice. Development 121:489–503

    Article  CAS  Google Scholar 

  • Louch WE, Sheehan KA, Wolska BM (2011) Methods in cardiomyocyte isolation, culture, and gene transfer. J Mol Cell Cardiol 51:288–298

    Article  CAS  Google Scholar 

  • Mehta J, Li D (1999) Inflammation in ischemic heart disease: response to tissue injury or a pathogenetic villain? Cardiovasc Res 43:291–299

    Article  CAS  Google Scholar 

  • Min J-Y, Sullivan MF, Yang Y, Zhang J-P, Converso KL, Morgan JP, Xiao Y-F (2002) Significant improvement of heart function by cotransplantation of human mesenchymal stem cells and fetal cardiomyocytes in postinfarcted pigs. Ann Thorac Surg 74:1568–1575

    Article  Google Scholar 

  • Mitcheson JS, Hancox JC, Levi AJ (1998) Cultured adult cardiac myocytes: future applications, culture methods, morphological and electrophysiological properties. Cardiovasc Res 39:280–300

    Article  CAS  Google Scholar 

  • Moscoso, I., Centeno, A., Lopez, E., Rodriguez-Barbosa, J., Santamarina, I., Filgueira, P., Sanchez, M., Dominguez-Perles, R., Penuelas-Rivas, G. & Domenech, N. Differentiation “in vitro” of primary and immortalized porcine mesenchymal stem cells into cardiomyocytes for cell transplantation. Transplantation proceedings, 2005. Elsevier, 481–482

  • Nakagawa O, Ogawa Y, Itoh H, Suga S-I, Komatsu Y, Kishimoto I, Nishino K, Yoshimasa T, Nakao K (1995) Rapid transcriptional activation and early mRNA turnover of brain natriuretic peptide in cardiocyte hypertrophy. Evidence for brain natriuretic peptide as an. J Clin Investig 96:1280–1287

    Article  CAS  Google Scholar 

  • Newman PJ, Newman DK (2003) Signal transduction pathways mediated by PECAM-1: new roles for an old molecule in platelet and vascular cell biology. Arterioscler Thromb Vasc Biol 23:953–964

    Article  CAS  Google Scholar 

  • Nursalim, A., Katili, P. A. & Santoso, T. 2016. Cellular cardiomyoplasty for myocardial infarction: A 2014 evidence-based update. Acta Medica Indonesiana, 46

  • Osborn L, Hession C, Tizard R, Vassallo C, Luhowskyj S, Chi-Rosso G, Lobb R (1989) Direct expression cloning of vascular cell adhesion molecule 1, a cytokine-induced endothelial protein that binds to lymphocytes. Cell 59:1203–1211

    Article  CAS  Google Scholar 

  • Piper H, Probst I, Schwartz P, Hütter F, Spieckermann P (1982) Culturing of calcium stable adult cardiac myocytes. J Mol Cell Cardiol 14:397–412

    Article  CAS  Google Scholar 

  • Pontén A, Walsh S, Malan D, Xian X, Schéele S, Tarnawski L, Fleischmann BK, Jovinge S (2013) FACS-based isolation, propagation and characterization of mouse embryonic cardiomyocytes based on VCAM-1 surface marker expression. PLoS One 8:e82403

    Article  Google Scholar 

  • Raeburn CD, Calkins CM, Zimmerman MA, Song Y, Ao L, Banerjee A, Harken AH, Meng X (2002) ICAM-1 and VCAM-1 mediate endotoxemic myocardial dysfunction independent of neutrophil accumulation. Am J Physiol-Regulatory, Integrat Comparative Physiol 283:R477–R486

    Article  CAS  Google Scholar 

  • Reedquist KA, Ross E, Koop EA, Wolthuis RM, Zwartkruis FJ, van Kooyk Y, Salmon M, Buckley CD, Bos JL (2000) The small GTPase, Rap1, mediates CD31-induced integrin adhesion. J Cell Biol 148:1151–1158

    Article  CAS  Google Scholar 

  • Reshef E, Sabbah HN, Nussinovitch U (2020) Effects of protective controlled coronary reperfusion on left ventricular remodeling in dogs with acute myocardial infarction: a pilot study. Cardiovasc Revasc Med 21:1579–1584

    Article  Google Scholar 

  • Sander V, Suñe G, Jopling C, Morera C, Belmonte JCI (2013) Isolation and in vitro culture of primary cardiomyocytes from adult zebrafish hearts. Nat Protoc 8:800

    Article  Google Scholar 

  • Schenkel AR, Chew TW, Muller WA (2004) Platelet endothelial cell adhesion molecule deficiency or blockade significantly reduces leukocyte emigration in a majority of mouse strains. J Immunol 173:6403–6408

    Article  CAS  Google Scholar 

  • Uosaki H, Fukushima H, Takeuchi A, Matsuoka S, Nakatsuji N, Yamanaka S, Yamashita JK (2011) Efficient and scalable purification of cardiomyocytes from human embryonic and induced pluripotent stem cells by VCAM1 surface expression. PloS one 6:e23657

    Article  CAS  Google Scholar 

  • Wab Naim WN, Mohamed Mokhtarudin MJ, Chan BT Lim, Ahmad Bakir E, Nik Mohamed A, NA. (2021) The study of myocardial ischemia-reperfusion treatment through computational modelling. J Theor Biol 509:110527

    Article  Google Scholar 

  • Zhou Y, Wang L, Liu Z, Alimohamadi S, Yin C, Liu J, Qian L (2017) Comparative gene expression analyses reveal distinct molecular signatures between differentially reprogrammed cardiomyocytes. Cell Rep 20:3014–3024

    Article  CAS  Google Scholar 

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Acknowledgements

The authors would like to thank the University of Indonesia for the grant supports No DLG-12/UN2.R3/PPM.00.00/2019, to all the donors that enable this study.

Funding

This study was supported by grant no. DLG-12/UN2.R3/PPM.00.00/2019 from the University of Indonesia.

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Authors and Affiliations

  1. Faculty of Medicine, Universitas Riau, Pekanbaru, 28293, Indonesia

    Normalina Sandora

  2. Indonesia Medical Education and Research Institute, Jakarta, 10430, Indonesia

    Normalina Sandora, Retno Wahyu Nurhayati,  Suwarti, Raisa Nauli, Tyas Rahmah Kusuma & Nur Amalina Fitria

  3. Faculty of Medicine, Universitas Indonesia, Jakarta, 10430, Indonesia

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

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  1. Normalina Sandora
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  2. Muhammad Arza Putra
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Correspondence to Normalina Sandora.

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The authors declare that there is no conflict of interest.

Ethical approval

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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Sandora, N., Putra, M.A., Nurhayati, R.W. et al. Characterisation of the single-cell human cardiomyocytes taken from the excess heart tissue of the right ventricular outlet in congenital heart disease. Cell Tissue Bank 23, 489–497 (2022). https://doi.org/10.1007/s10561-021-09970-4

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  • DOI: https://doi.org/10.1007/s10561-021-09970-4

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