A Method for Obtaining Polymorphonuclear Leukocytes from Intraperitoneal Exudates

• Published in: Blood 1958-12, Vol.13 (12), p.1192-1197
• Main Authors: Estes, Frances L., Smith, Shirley, Gast, Joseph H.
• Format: Article
• Language: English
• Subjects: Exudates and Transudates; Humans; Leukocytes; Leukocytosis; Neutrophils; Old Medline
• ISSN: 0006-4971; 1528-0020
• DOI: 10.1182/blood.V13.12.1192.1192

THE IN VITRO STUDY of normal leukocyte metabolism has been complicated by the presence of erythrocytes and platelets in the cell suspensions and by the need for a good yield of cells of reasonably constant characteristics. Because of the difficulties involved in recovering uncontaminated leukocytes of a single type from blood, a study of the method for obtaining the cells from intraperitoneal exudates was undertaken. The original method for obtaining leukocytes from intraperitoneal exudates was that of Hamburger.1 As used by Mudd et al.,2 the production of polymorphonuclear leukocytes was stimulated by an injection of 200 ml. of 0.9 per cent NaCl. The mixture of cells and fluid was recovered from three to four hours later by puncturing the cavity. Clotting was prevented by mixing the recovered fluid with sodium citrate. Barron and Harrop3 found that from rabbits the recovered cells gave inconsistent results in metabolic studies. They attributed the loss of oxidative capacity of the cells and high glycolytic quotient in aerobic glycolysis to damage from centrifugation and the presence of sodium citrate. Recently, Stahelin, Suter and Karnovsky4 examined exudates from guinea pigs and found a linear relationship between the rate of respiration and the relative number of polymorphonuclear leukocytes to monocytes. The presence of serum in the suspending medium has been reported both to increase5 and not to affect4 the respiration. Thus in order to study the metabolism of polymorphonuclear leukocytes it is highly desirable that the cell suspensions be uncontaminated by other types of cells and be free from clumps. Likewise, the suspending medium should be carefully buffered but free from extraneous material. With guinea pigs as the test animal a technic has been developed for obtaining from 80 to 90 per cent polymorphonuclear leukocytes from the intraperitoneal exudate. The recovered cells were from 91 to 98 per cent viable at the time of harvest, as indicated by vital staining. The oxygen consumption of individual batches of cells was reproducible within the limitations of the methods used. Preliminary experiments indicated that this technic can be used with rats as well.