Preservation of Human Granulocytes. II. Characteristics of Granulocytes Obtained by Counterflow Centrifugation.
Abstract
Human granulocytes were isolated from whole blood and from the buffy coat of blood by counterflow centrifugation using the Beckman JE-6 rotor. Ninety-one percent of granulocytes in the human blood sample were isolated in 59 studies. The differential white blood cell count showed 97% Polys, 1% lymphs, and 2% others. Measurements of oxygen consumption, chemotaxis, and phagocytosis in granulocytes isolated by counterflow centrifugation were similar to those in granulocytes isolated from blood by dextran sedimentation. The granulocytes had normal adenine and guanine di-and trinucleotide levels. After fluorescein diacetate treatment, the cytoplasm of 98.5 + or - 0.7% of fresh granulocytes was positive, while 98.5 + or - 1.2% of the fresh granulocytes excluded ethidium bromide from their nuclei. The response of freeze-preserved granulocytes to treatment with fluorescein diacetate and ethidium bromide showed that 75% of granulocytes survived freezing and thawing in 5% DMSO but were progressively unstable after incubation for 2 hours at 37C.
Document Details
- Document Type
- Technical Report
- Publication Date
- Oct 25, 1976
- Accession Number
- ADA031634
Entities
People
- C. R. Valeri
- Fabian J. Lionetti
- P. S. Lin
- S. R. Kurtz
- Stephen M. Hunt