Cell-type specificity of preconditioning in an in vitro model

Y. Liu, W. D. Gao, B. O'Rourke, E. Marban

Research output: Contribution to journalArticle

Abstract

We investigated whether preconditioning could protect several cultured cell lines, to determine whether the protection is specific for cells derived from different myogenic and non-myogenic sources. Ischemia was simulated by centrifugation of cells into a pellet, and cell viability was determined by hypotonic trypan blue solution. Preconditioning was produced by brief exposures to either glucose-free solution or metabolic inhibition. Freshly isolated rabbit ventricular myocytes were studied to confirm that preconditioning occurs in this model. We then compared these results to those in several cultured cell lines, including HEK 293 cells derived from human embryonic kidney, HIT-T15 cells from Syrian hamster pancreatic islets, and C2C12 cells from mouse skeletal muscle. In the latter cell line, we also determined whether differentiation alters preconditioning. Preconditioning protected rabbit ventricular myocytes: the percentage of dead cells was decreased from 36.8 ± 4.7% in the control group to 23.0 ± 5.2% in the preconditioned group after 60 min and from 50.7 ± 2.1% in the control group to 25.5 ± 4.5% in the preconditioned group after 120 min ischemia (p < 0.02). In contrast, there was no protection from preconditioning in HEK 293 cells or HIT-T15 cells. Preconditioning did not protect C2C12 myoblasts either. Interestingly, after C2C12 myoblasts had differentiated into myotubes (induced by exposing the cells to low-serum medium), they could then be protected by preconditioning (46.3 ± 3.6% in the control group vs 26.0 ± 2.7% in the preconditioned group after 60 min and 67.4 ± 3.6% in the control group vs 46.0 ± 4.6% in the preconditioned group after 120 min ischemia; p < 0.05). In conclusion, protection from preconditioning is cell-type specific. The presence of endogenous K(ATP) channels (which are plentiful in HIT-T15 cells) is insufficient to enable preconditioning of the cell. Among the various cell types studied, only differentiated muscle cells (rabbit ventricular myocytes and C2C12 myotubes) exhibited preconditioning.

Original languageEnglish (US)
Pages (from-to)450-457
Number of pages8
JournalBasic Research in Cardiology
Volume91
Issue number6
DOIs
StatePublished - Nov 1996

Keywords

  • CC cell line
  • Differentiation
  • HEK 293 cell line
  • HIT-T15 cell line
  • Ischemic preconditioning
  • Rabbit ventricular myocytes

ASJC Scopus subject areas

  • Physiology
  • Cardiology and Cardiovascular Medicine
  • Physiology (medical)

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