Cold preservation of endothelial cells in sucrose-based solution (SbS) and University of Wisconsin (UW) solutions: comparison of normoxic or hypoxic storage.
Cold preservation of endothelial cells was studied, comparing primary endothelial cells (human umbilical vein endothelial cells - HUVECs) and a continuously growing cell line (ECV304 cells). Viability at the end of 24h cold preservation was measured by dye exclusion, whilst metabolism was assessed by Alamar blue conversion. Two preservation solutions were studied (UW solution) and sucrose-based (SbS) in both cell types. The response was similar in both cell types to preservation under normoxic conditions (with percentage dye exclusion maintained at about 80 percent in both preservation solutions) whereas under hypoxic conditions ECV304 were more sensitive to preservation in UW solution (dye exclusion reduced to 43.5+/-1.4 percent versus 73.6+/-14 percent (P<0.01). Metabolism assessed by Alamar blue conversion after cold preservation and rewarming was similar in both ECV304 and HUVECs after storage under normoxic conditions in UW solution, but in both cell types, metabolism was higher in SbS (P<0.05 and p<0.01) than in UW solution. Under hypoxic conditions, both cell types showed similar recovery of metabolism after storage in either UW or SbS. If the cells (in this case ECV304 under aerobic conditions) were stored for 24h and then allowed to rewarm in either of the respective preservation solutions (UW or SbS for 1h) before the Alamar blue test, metabolism was higher (p less than 0.01) in those exposed to SbS. UW solution and SbS provide similar protection for endothelial cells under hypoxic conditions, but SbS has some advantages under normoxic storage or if the cells experience variable temperatures in the presence of residual preservation solution at the end of cold preservation period.
University Department of Surgery, Royal Free Hospital and University College Medical School, London NW3 2QG, UK.
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M
Hawkins
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Sales
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Dijk
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Fuller
D000241:Adenosine; D000493:Allopurinol; D015687:Cell Hypoxia; D002460:Cell Line; D002470:Cell Survival; D002478:Cells, Cultured; D015925:Cryopreservation; D003451:Cryoprotective Agents; D042783:Endothelial Cells / Q000166:cytology*; D005978:Glutathione; D006801:Humans; D007328:Insulin; D019188:Organ Preservation Solutions; D011887:Raffinose; D013395:Sucrose; D014471:Umbilical Veins
D003451:Cryoprotective Agents; D007328:Insulin; D019188:Organ Preservation Solutions; C055206:University of Wisconsin-lactobionate solution; D013395:Sucrose; D000493:Allopurinol; D005978:Glutathione; D000241:Adenosine; D011887:Raffinose