Human lung fibroblasts stored in preservation solutions containing trehalose: Does it increase cell viability?

Paolo Carbognani, M. Rusca, L. Spaggiari, L. Cattelani, P. Solli, P. Bobbio

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Abstract

Purpose: The research on lung preservation for transplantation is directed to synthetize a solution that avoids the damages of lung tissues caused by ischemia. Trehalose is a non-reducing disaccharide that stabilizes cell membrane under various stressing conditions. In order to verify its presumed cytoprotective effects we have evaluated human lung fibroblasts viability after storage in EuroCollins (EC), Low Potassium Dextran (LPD) and RingerLactate (RL) solutions, in which trehalose has replaced glucose (T-EC) or was added (LPD-T, RL-T). Methods: Normal human diploid fibroblasts derived from foetal lung (WI-38) after storage in Basal Eagle's medium, were seeded with an average density of 10,000 cells/cm2 in 9cm2 well plates. Subcultures were incubated at 10°C for 12 hours in standard and modified EC, LPD and RL. Then the cells were put in growth medium containing 40 μM of Leucine and supplied with labelled Leucine (2 μCi/ml) for 30 minutes. The modified EC solution was obtained replacing glucose by 35.0 gm/L (3.5%) of trehalose reaching an osmolarity of 295 mosM; this disaccharide at the same concentration was added to LPD and to RL with respectively a final osmolarity of 350 mosM and 325 mosM. Cell viability has been evaluated on the basis of the rate of protein synthesis expressed as nMol of 3H Leucine/mg of proteins/30 minutes. The data related to 3 wells for each tested solution are presented as mean ± standard deviation and evaluated with the Student's t-test. Results: After 12 hours of incubation in standard and modified solutions the data are as follows. At time O the Control Value 3.195±0.429; EC 0.134±0.021, T-EC 0.125±0.007 (p>0.05); LPD 0.561±0.28, LPD-T 0.160±0.007 (p

Original languageEnglish
JournalChest
Volume110
Issue number4 SUPPL.
Publication statusPublished - Oct 1996

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ASJC Scopus subject areas

  • Pulmonary and Respiratory Medicine

Cite this

Carbognani, P., Rusca, M., Spaggiari, L., Cattelani, L., Solli, P., & Bobbio, P. (1996). Human lung fibroblasts stored in preservation solutions containing trehalose: Does it increase cell viability? Chest, 110(4 SUPPL.).