Annals of Biomedical Engineering

, Volume 35, Issue 8, pp 1414–1424 | Cite as

An In Vitro System to Evaluate the Effects of Ischemia on Survival of Cells Used for Cell Therapy

  • Bryce H. Davis
  • Thies Schroeder
  • Pavel S. Yarmolenko
  • Farshid Guilak
  • Mark W. Dewhirst
  • Doris A. Taylor
Article

Abstract

Maintaining cell viability is a major challenge associated with transplanting cells into ischemic myocardium to restore function. A likely contributor to significant cell death during cardiac cell therapy is hypoxia/anoxia. We developed a system that enabled quantification and association of cell survival with oxygen and nutrient values within in vitro constructs. Myoblasts were suspended in 2% collagen gels in 1 cm diameter × 1 cm deep constructs. At 48 ± 3 h post-seeding, oxygen levels were measured using microelectrodes and gels were snap-frozen. Bioluminescence metabolite imaging and TUNEL staining were performed on cryosections. Oxygen and glucose consumption and lactate production rates were calculated by fitting data to Fick’s second law of diffusion with Michaelis–Menten kinetics. Oxygen levels dropped to 0 mmHg and glucose levels dropped from 4.28 to 3.18 mM within the first 2000 μm of construct depth. Cell viability dropped to approximately 40% over that same distance and continued to drop further into the construct. We believe this system provides a reproducible and controllable test bed to compare survival, proliferation, and phenotype of various cell inputs (e.g., myoblasts, mesenchymal stem cells, and cardiac stem cells) and the impact of different treatment regimens on the likelihood of survival of transplanted cells.

Keywords

Myoblast Ischemia Stem cell Cardiomyoplasty Myocardial infarction 

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Copyright information

© Biomedical Engineering Society 2007

Authors and Affiliations

  • Bryce H. Davis
    • 1
  • Thies Schroeder
    • 2
  • Pavel S. Yarmolenko
    • 2
  • Farshid Guilak
    • 1
    • 3
  • Mark W. Dewhirst
    • 2
  • Doris A. Taylor
    • 1
    • 4
  1. 1.Department of Biomedical EngineeringDuke UniversityDurhamUSA
  2. 2.Department of Radiation OncologyDuke University Medical Center DurhamUSA
  3. 3.Department of Surgery, Division of Orthopedic SurgeryDuke University Medical Center DurhamUSA
  4. 4.Center for Cardiovascular Repair, Department of Physiology, College of MedicineUniversity of MinnesotaMinneapolisUSA

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