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

Normalina Sandora, Muhammad Arza Putra, Retno Wahyu Nurhayati, Suwarti, Raisa Nauli, Tyas Rahmah Kusuma, Nur Amalina Fitria, Ardiansyah, Chaidar Muttaqin, William Makdinata, Idrus Alwi

Research output: Contribution to journalArticlepeer-review

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.

Original languageEnglish
JournalCell and Tissue Banking
DOIs
Publication statusAccepted/In press - 2021

Keywords

  • Cardiac progenitor cells
  • Cardiomyocyte isolation
  • Right ventricular resection
  • Tetralogy of fallot
  • Waste heart tissue

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