Document Detail

Validation of a new live cell strain system: characterization of plasma membrane stress failure.
MedLine Citation:
PMID:  11356803     Owner:  NLM     Status:  MEDLINE    
Motivated by our interest in lung deformation injury, we report on the validation of a new live cell strain system. We showed that the system maintains a cell culture environment equivalent to that provided by conventional incubators and that its strain ouput was uniform and reproducible. With this system, we defined cell deformation dose (i.e., membrane strain amplitude)-cell injury response relationships in alveolar epithelial cultures and studied the effects of temperature on them. Deformation injury occurred in the form of reversible, nonlethal plasma membrane stress failure events and was quantified as the fraction of cells with uptake and retention of fluorescein-labeled dextran (FITC-Dx). The undeformed control population showed virtually no FITC-Dx uptake at any temperature, which was also true for cells strained by 3%. However, when the membrane strain was increased to 18%, ~5% of cells experienced deformation injury at a temperature of 37 degrees C. Moreover, at that strain, a reduction in temperature to 4 degrees C resulted in a threefold increase in the number of cells with plasma membrane breaks (from 4.8 to 15.9%; P < 0.05). Cooling of cells to 4 degrees C also lowered the strain threshold at which deformation injury was first seen. That is, at a 9% substratum strain, cooling to 4 degrees C resulted in a 10-fold increase in the number of cells with FITC-Dx staining (0.7 vs. 7.5%, P < 0.05). At that temperature, A549 cells offered a 50% higher resistance to shape change (magnetic twisting cytometry measurements) than at 37 degrees C. We conclude that the strain-injury threshold of A549 cells is reduced at low temperatures, and we consider temperature effects on plasma-membrane fluidity, cytoskeletal stiffness, and lipid trafficking as responsible mechanisms.
R W Stroetz; N E Vlahakis; B J Walters; M A Schroeder; R D Hubmayr
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S.; Validation Studies    
Journal Detail:
Title:  Journal of applied physiology (Bethesda, Md. : 1985)     Volume:  90     ISSN:  8750-7587     ISO Abbreviation:  J. Appl. Physiol.     Publication Date:  2001 Jun 
Date Detail:
Created Date:  2001-05-17     Completed Date:  2001-07-26     Revised Date:  2013-09-26    
Medline Journal Info:
Nlm Unique ID:  8502536     Medline TA:  J Appl Physiol (1985)     Country:  United States    
Other Details:
Languages:  eng     Pagination:  2361-70     Citation Subset:  IM    
Thoracic Diseases Research Unit, Division of Pulmonary and Critical Care Medicine, Department of Internal Medicine, Mayo Clinic and Foundation, Rochester, Minnesota 55905, USA.
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MeSH Terms
Cell Division / physiology
Cell Line
Cell Membrane / physiology*,  ultrastructure
Cell Survival
Epithelial Cells / physiology*
Flow Cytometry
Models, Biological
Pulmonary Alveoli / cytology,  physiology*
Respiration, Artificial*
Stress, Mechanical*
Grant Support

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