A determination of biophysical parameters related to freezing of an ELT-3 cell line

Marwane S. Berrada, John C. Bischof

Research output: Chapter in Book/Report/Conference proceedingConference contribution

7 Scopus citations

Abstract

This study investigates two destructive biophysicalmechanisms during freezing (extensive dehydration andintracellular ice formation) at the cellular level in the rodentELT-3 uterine fibroid cell-line. The osmotically inactivevolume fraction (Vb) of ELT-3 cells was approximated to 0.35of the initial isotonic cell volume (V0). The water transportcharacteristics of this cell-line are such that ELT-3 cells arehighly permeable with a strong ability to lose water even at lowsubzero temperatures. The hydraulic reference permeability, Lpgand activation energy, Eip associated with Lp were found to be0.13 (jim/min.atm) and 19.0 (kcal/mole) [R2=0.86]respectively. Intracellular Ice Formation (IIF) occurs at lowertemperatures than many cell-types (i.e. TnF 50% below -15°C)at cooling rates > 25 °C/min. Darkening IIF, which wasassumed to occur by Surface Catalyzed Nucleation (SCN), isgoverned by kinetic Q0 and thermodynamic K0 biophysicalparameters, which were found to be 6.1xl08(m2.s)'' and5.3xl09(K5) [R2=0.94] respectively. At a cooling rate of100°C/min, twitching IIF (non-darkening IIF) was observed.Viability data from a separate study (Bischof et. al., 2000)indicated that at cooling rates < l°C/min and > 50°C/min withan enderature of -20°C, extensive damage to cells wasobserved. The current biophysical study shows that extensivedehydration occurs at l°C/min while substantial IIF (77%)occurs at 50 °C/min. This data suggests that while biophysicscan explain some of the destruction occurring at theinvestigated temperatures, other effects or mechanisms may beplaying a role at lower enderatures.

Original languageEnglish (US)
Title of host publicationAdvances in Heat and Mass Transfer in Biotechnology
PublisherAmerican Society of Mechanical Engineers (ASME)
Pages41-48
Number of pages8
ISBN (Electronic)9780791819296
DOIs
StatePublished - 2000
EventASME 2000 International Mechanical Engineering Congress and Exposition, IMECE 2000 - Orlando, United States
Duration: Nov 5 2000Nov 10 2000

Publication series

NameASME International Mechanical Engineering Congress and Exposition, Proceedings (IMECE)
Volume2000-F

Conference

ConferenceASME 2000 International Mechanical Engineering Congress and Exposition, IMECE 2000
Country/TerritoryUnited States
CityOrlando
Period11/5/0011/10/00

Bibliographical note

Funding Information:
The ELT-3 cell line was a gift to the Cell Injury Laboratory (University of Minnesota, Minneapolis, MN) from Dr. Cheryl Walker, MD (Anderson Cancer Center, Smithville, TX).

Publisher Copyright:
© 2000 American Society of Mechanical Engineers (ASME). All rights reserved.

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