Isolation and Cryogenic Preservation of Monocytes from Plateletpheresis Cellular Residues.
Abstract
Human monocytes were isolated from the cellular residues afte plateletpheresis of donars using the Haemonetics Model 30 Blood Cell Processor. Mononuclear cells were obtained with Ficoll-Isopaque and separated from lymphocytes by stepwise elutriation in a Beckman JE-6 rotor equipped with two isolation chambers. The isolated cells were frozen in a solution containing an extracellular (dimethylsulfoxide, DMSO) cryoprotective compound. In three procedures, approximately 1 x 10 to the 9th power monocytes were obtained. Ninety-nine percent of isolated monocytes were viable in the fluorescein diacetate (FDA)-ethidium bromide (EB) test. Myeloperoxidase-positive cells were 95% and 90% respectively in Chambers nos. 1 and 2. Ninety-four percent of monocytes ingested 5 or more opsonized Fluolite particles and 95% ingested 1 or more ethidium-treated zymosan particles. After storage in liquid nitrogen for up to 9 weeks, 99% of the cells were recovered after thawing. Of these, 95% were myeloperoxidase-positive, 94% showed intact memranes in the FDA-EB test, 95% ingested 5 or more opsonized Fluolite particles, an 96% ingested 1 or more ethidium-treated xymosan particles.
Document Details
- Document Type
- Technical Report
- Publication Date
- Jan 11, 1983
- Accession Number
- ADA123675
Entities
People
- C. Robert Valeri
- Fabian J. Lionetti
- Stephen M. Hunt