Neutrophil migration in canines: in vivo comparison of granulocyte function following 24-hour storage at 6 or 20 degrees C of leukocyte concentrates or of granulocytes purified by counterflow centrifugation--elutriation.
The use of granulocyte-rich concentrates from leukaphresis purified by counterflow centrifugation--elutriation to obtain pure granulocytes for transfusion studies in cyclophosphamide-induced neutropenic animal models is reported. Our data for granulocyte-rich leukapheresis concentrates indicate that room temperature (20 degrees C) appears to be preferred to 6 degrees C for short-term granulocyte storage. The data also indicate that although the granulocytes isolated by counterflow centrifugation--elutration may retain in vitro functions of chemotaxis, phagocytosis, and bactericidal activity, the in vivo function of migration into skin chambers for isolated granulocytes is seriously impaired after storage for 18 to 24 hr at both 6 and 20 degrees C. This loss of in vivo function of stored granulocytes occurs in isolated granulocytes obtained by both counterflow centrifugation--elutriation and dextran sedimentation, and it is not observed in the leukocyte concentrates held at 20 degrees C. The results of these studies are four-fold. First, freshly isolated granulocytes display no apparent loss of either in vivo or in vitro function. Second, granulocytes isolated by counterflow centrifugation--elutriation or dextran sedimentation and stored at 6 or 20 degrees C are severely impaired in terms of their in vivo chemotactic function but display no loss of in vitro efficacy. Third, 20 degrees C storage of granulocyte-rich leukapheresis concentrates for 18 to 24 hr is superior to 6 degrees C storage. Fourth, in vitro analysis may be limited in its ability to indicate in vivo function as a measure of success in granulocyte preservation studies.
D000818:Animals; D001793:Blood Preservation; D001803:Blood Transfusion; D002469:Cell Separation / Q000379:methods; D002470:Cell Survival; D002634:Chemotaxis, Leukocyte; D003080:Cold Temperature; D004285:Dogs; D005260:Female; D006098:Granulocytes / Q000502:physiology* / Q000637:transplantation; D066298:In Vitro Techniques; D007962:Leukocytes / Q000502:physiology; D008297:Male; D009504:Neutrophils / Q000502:physiology; D013997:Time Factors