The effect of low-potassium-dextran versus Euro-Collins solution for preservation of isolated type II pneumocytes.
Limited availability of donor organs is a major factor restricting the clinical application of lung transplantation. Improvements in preservation techniques are essential for prolonging storage time and improving lung function following transplantation. The present investigation used primary cultures of adult rat alveolar type II cells as a model for evaluating lung-preservation solutions. Type II cells were plated onto tissue-culture plastic at a density 5 x 10(5) cells/cm2 and maintained in Dulbecco's modified Eagle's medium containing 10% fetal bovine serum (D10) for 40 hr. Cells were then exposed to Euro-Collins solution or a low-potassium-dextran solution (LPD). At designated time points, measurements of lactate-dehydrogenase (LDH) release, protein content, and incorporation of 3H-thymidine into cellular DNA were made. During 12 hr of "storage" at 37 degrees C, cells maintained in LPD released less LDH (14.3 +/- 1.2% of cellular total, mean +/- SEM, n = 5) than their counterparts stored in EC (20.6 +/- 1.6%, P less than 0.05). During the 36 hr following a 6-hr exposure to preservative solutions, LPD-treated cells incorporated more thymidine per mg of protein (2566 +/- 419.8 cpm/micrograms protein, mean +/- SEM, n = 6) compared with cells maintained continuously in D10 (1431 +/- 351, P less than 0.05). By contrast, cells exposed to EC incorporated less thymidine (82.2 +/- 62.8 cpm/micrograms protein) than either cells maintained in LPD or D10 (P less than 0.01 for each comparison). These results suggest that LPD solution is less cytotoxic than EC and that LPD enables higher levels of metabolic activity in recovering epithelial cells. In vitro cultures of type II epithelial cells are a useful model system for the study of lung preservation and posttransplantation lung injury.
Maccherini
M
M
Department of Medicine, Toronto Hospital, Mt. Sinai Hospital, Canada.
Keshavjee
S H
SH
Slutsky
A S
AS
Patterson
G A
GA
Edelson
J D
JD
D000469:Alkaline Phosphatase; D000818:Animals; D002478:Cells, Cultured; D003911:Dextrans; D004847:Epithelial Cells; D004848:Epithelium / Q000187:drug effects; D006863:Hydrogen-Ion Concentration; D006982:Hypertonic Solutions; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008168:Lung / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D008297:Male; D009926:Organ Preservation / Q000379:methods*; D011188:Potassium; D011650:Pulmonary Alveoli / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D051381:Rats; D011919:Rats, Inbred Strains; D012996:Solutions
D003911:Dextrans; C037096:Euro-Collins' solution; D006982:Hypertonic Solutions; D012996:Solutions; D007770:L-Lactate Dehydrogenase; D000469:Alkaline Phosphatase; D011188:Potassium