A novel cell culture model for studying ischemia-reperfusion injury in lung transplantation

Jonathan A. Cardella, Shaf Keshavjee, Eric Mourgeon, Stephen D. Cassivi, Stefan Fischer, Noritaka Isowa, Arthur Slutsky, Mingyao Liu

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

Many cell culture models have been developed to study ischemia-reperfusion injury; however, none is specific to the conditions of lung preservation and transplantation. The objective of this study was to design a cell culture model that mimics clinical lung transplantation, in which preservation is aerobic and hypothermic. A549 cells, a human pulmonary epithelial cell line, were preserved in 100% O2 at 4°C for varying periods in low-potassium dextran glucose solution, simulating ischemia, followed by the introduction of warm (37°C) DMEM plus 10% fetal bovine serum to simulate reperfusion. Cultures were assayed for cell attachment and viability. Sequential extension of ischemic times to 24 h showed a time-dependent loss of cells. There was a further decrease in cell number after simulated reperfusion. Cell detachment was due mainly to cell death, as determined by cell viability. The effects of chemical components such as dextran 40 and calcium in the preservation solution and various preservation gas mixtures were examined by use of this model system. With its design and validation, this model could be used to study mechanisms related to ischemia-reperfusion injury at the cellular and molecular level.

Original languageEnglish (US)
Pages (from-to)1553-1560
Number of pages8
JournalJournal of Applied Physiology
Volume89
Issue number4
StatePublished - 2000
Externally publishedYes

Fingerprint

Lung Transplantation
Reperfusion Injury
Reperfusion
Cell Survival
Cell Culture Techniques
Dextrans
Cell Death
Ischemia
Cell Count
Gases
Epithelial Cells
Calcium
Cell Line
Lung
Serum
A549 Cells
low potassium dextran glucose solution

Keywords

  • Acute lung injury
  • Cell viability
  • Epithelial cells
  • Organ preservation

ASJC Scopus subject areas

  • Physiology
  • Endocrinology
  • Orthopedics and Sports Medicine
  • Physical Therapy, Sports Therapy and Rehabilitation

Cite this

Cardella, J. A., Keshavjee, S., Mourgeon, E., Cassivi, S. D., Fischer, S., Isowa, N., ... Liu, M. (2000). A novel cell culture model for studying ischemia-reperfusion injury in lung transplantation. Journal of Applied Physiology, 89(4), 1553-1560.

A novel cell culture model for studying ischemia-reperfusion injury in lung transplantation. / Cardella, Jonathan A.; Keshavjee, Shaf; Mourgeon, Eric; Cassivi, Stephen D.; Fischer, Stefan; Isowa, Noritaka; Slutsky, Arthur; Liu, Mingyao.

In: Journal of Applied Physiology, Vol. 89, No. 4, 2000, p. 1553-1560.

Research output: Contribution to journalArticle

Cardella, JA, Keshavjee, S, Mourgeon, E, Cassivi, SD, Fischer, S, Isowa, N, Slutsky, A & Liu, M 2000, 'A novel cell culture model for studying ischemia-reperfusion injury in lung transplantation', Journal of Applied Physiology, vol. 89, no. 4, pp. 1553-1560.
Cardella JA, Keshavjee S, Mourgeon E, Cassivi SD, Fischer S, Isowa N et al. A novel cell culture model for studying ischemia-reperfusion injury in lung transplantation. Journal of Applied Physiology. 2000;89(4):1553-1560.
Cardella, Jonathan A. ; Keshavjee, Shaf ; Mourgeon, Eric ; Cassivi, Stephen D. ; Fischer, Stefan ; Isowa, Noritaka ; Slutsky, Arthur ; Liu, Mingyao. / A novel cell culture model for studying ischemia-reperfusion injury in lung transplantation. In: Journal of Applied Physiology. 2000 ; Vol. 89, No. 4. pp. 1553-1560.
@article{ae8c7976b5084d39943dc21e9b845a91,
title = "A novel cell culture model for studying ischemia-reperfusion injury in lung transplantation",
abstract = "Many cell culture models have been developed to study ischemia-reperfusion injury; however, none is specific to the conditions of lung preservation and transplantation. The objective of this study was to design a cell culture model that mimics clinical lung transplantation, in which preservation is aerobic and hypothermic. A549 cells, a human pulmonary epithelial cell line, were preserved in 100{\%} O2 at 4°C for varying periods in low-potassium dextran glucose solution, simulating ischemia, followed by the introduction of warm (37°C) DMEM plus 10{\%} fetal bovine serum to simulate reperfusion. Cultures were assayed for cell attachment and viability. Sequential extension of ischemic times to 24 h showed a time-dependent loss of cells. There was a further decrease in cell number after simulated reperfusion. Cell detachment was due mainly to cell death, as determined by cell viability. The effects of chemical components such as dextran 40 and calcium in the preservation solution and various preservation gas mixtures were examined by use of this model system. With its design and validation, this model could be used to study mechanisms related to ischemia-reperfusion injury at the cellular and molecular level.",
keywords = "Acute lung injury, Cell viability, Epithelial cells, Organ preservation",
author = "Cardella, {Jonathan A.} and Shaf Keshavjee and Eric Mourgeon and Cassivi, {Stephen D.} and Stefan Fischer and Noritaka Isowa and Arthur Slutsky and Mingyao Liu",
year = "2000",
language = "English (US)",
volume = "89",
pages = "1553--1560",
journal = "Journal of Applied Physiology",
issn = "8750-7587",
publisher = "American Physiological Society",
number = "4",

}

TY - JOUR

T1 - A novel cell culture model for studying ischemia-reperfusion injury in lung transplantation

AU - Cardella, Jonathan A.

AU - Keshavjee, Shaf

AU - Mourgeon, Eric

AU - Cassivi, Stephen D.

AU - Fischer, Stefan

AU - Isowa, Noritaka

AU - Slutsky, Arthur

AU - Liu, Mingyao

PY - 2000

Y1 - 2000

N2 - Many cell culture models have been developed to study ischemia-reperfusion injury; however, none is specific to the conditions of lung preservation and transplantation. The objective of this study was to design a cell culture model that mimics clinical lung transplantation, in which preservation is aerobic and hypothermic. A549 cells, a human pulmonary epithelial cell line, were preserved in 100% O2 at 4°C for varying periods in low-potassium dextran glucose solution, simulating ischemia, followed by the introduction of warm (37°C) DMEM plus 10% fetal bovine serum to simulate reperfusion. Cultures were assayed for cell attachment and viability. Sequential extension of ischemic times to 24 h showed a time-dependent loss of cells. There was a further decrease in cell number after simulated reperfusion. Cell detachment was due mainly to cell death, as determined by cell viability. The effects of chemical components such as dextran 40 and calcium in the preservation solution and various preservation gas mixtures were examined by use of this model system. With its design and validation, this model could be used to study mechanisms related to ischemia-reperfusion injury at the cellular and molecular level.

AB - Many cell culture models have been developed to study ischemia-reperfusion injury; however, none is specific to the conditions of lung preservation and transplantation. The objective of this study was to design a cell culture model that mimics clinical lung transplantation, in which preservation is aerobic and hypothermic. A549 cells, a human pulmonary epithelial cell line, were preserved in 100% O2 at 4°C for varying periods in low-potassium dextran glucose solution, simulating ischemia, followed by the introduction of warm (37°C) DMEM plus 10% fetal bovine serum to simulate reperfusion. Cultures were assayed for cell attachment and viability. Sequential extension of ischemic times to 24 h showed a time-dependent loss of cells. There was a further decrease in cell number after simulated reperfusion. Cell detachment was due mainly to cell death, as determined by cell viability. The effects of chemical components such as dextran 40 and calcium in the preservation solution and various preservation gas mixtures were examined by use of this model system. With its design and validation, this model could be used to study mechanisms related to ischemia-reperfusion injury at the cellular and molecular level.

KW - Acute lung injury

KW - Cell viability

KW - Epithelial cells

KW - Organ preservation

UR - http://www.scopus.com/inward/record.url?scp=0033774822&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0033774822&partnerID=8YFLogxK

M3 - Article

C2 - 11007595

AN - SCOPUS:0033774822

VL - 89

SP - 1553

EP - 1560

JO - Journal of Applied Physiology

JF - Journal of Applied Physiology

SN - 8750-7587

IS - 4

ER -