Development of an In Vitro Cardiac Ischemic Model Using Primary Human Cardiomyocytes

Pezhman Hafez, Shiplu Roy Chowdhury, Shinsmon Jose, Jia Xian Law, Ruszymah Idrus, Mohd Ramzisham Abdul Rahman, Min Hwei Ng

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Developing experimental models to study ischemic heart disease is necessary for understanding of biological mechanisms to improve the therapeutic approaches for restoring cardiomyocytes function following injury. The aim of this study was to develop an in vitro hypoxic/re-oxygenation model of ischemia using primary human cardiomyocytes (HCM) and define subsequent cytotoxic effects. HCM were cultured in serum and glucose free medium in hypoxic condition with 1% O2 ranging from 30 min to 12 h. The optimal hypoxic exposure time was determined using Hypoxia Inducible Factor 1α (HIF-1α) as the hypoxic marker. Subsequently, the cells were moved to normoxic condition for 3, 6 and 9 h to replicate the re-oxygenation phase. Optimal period of hypoxic/re-oxygenation was determined based on 50% mitochondrial injury via 3-(4,5-dimethylthiazol-2-Yl)-2,5-diphenyltetrazolium bromide assay and cytotoxicity via lactate dehydrogenase (LDH) assay. It was found that the number of cells expressing HIF-1α increased with hypoxic time and 3 h was sufficient to stimulate the expression of this marker in all the cells. Upon re-oxygenation, mitochondrial activity reduced significantly whereas the cytotoxicity increased significantly with time. Six hours of re-oxygenation was optimal to induce reversible cell injury. The injury became irreversible after 9 h as indicated by > 60% LDH leakage compared to the control group cultured in normal condition. Under optimized hypoxic reoxygenation experimental conditions, mesenchymal stem cells formed nanotube with ischemic HCM and facilitated transfer of mitochondria suggesting the feasibility of using this as a model system to study molecular mechanisms of myocardial injury and rescue.

Original languageEnglish
Pages (from-to)529-538
Number of pages10
JournalCardiovascular Engineering and Technology
Volume9
Issue number3
DOIs
Publication statusPublished - 15 Sep 2018

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Oxygenation
Cardiac Myocytes
Wounds and Injuries
Hypoxia-Inducible Factor 1
Cytotoxicity
L-Lactate Dehydrogenase
Assays
Nanotubes
Cell Hypoxia
Mitochondria
Serum-Free Culture Media
Stem cells
Mesenchymal Stromal Cells
Myocardial Ischemia
Glucose
Theoretical Models
Ischemia
Cells
In Vitro Techniques
Control Groups

Keywords

  • Cardiac ischemic model
  • Hypoxia/re-oxygenation
  • Mitochondrial transfer
  • Primary human cardiomyocytes
  • Reperfusion injury

ASJC Scopus subject areas

  • Biomedical Engineering
  • Cardiology and Cardiovascular Medicine

Cite this

Development of an In Vitro Cardiac Ischemic Model Using Primary Human Cardiomyocytes. / Hafez, Pezhman; Chowdhury, Shiplu Roy; Jose, Shinsmon; Law, Jia Xian; Idrus, Ruszymah; Abdul Rahman, Mohd Ramzisham; Ng, Min Hwei.

In: Cardiovascular Engineering and Technology, Vol. 9, No. 3, 15.09.2018, p. 529-538.

Research output: Contribution to journalArticle

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