Collection, Separation, Cryopreservation and Characterization of Peripheral Blood and Bone Marrow Stem Cells and Their Use in Treating Lethally Irradiated Dogs.
Abstract
Peripheral blood mononuclear cells (MNC) were obtained from the buffy coats collected during 21 platelet aphersis procedures using the Haemonetics 30 Blood Processor. The mononuclear cells in the buffy coat were divided into two equal portions. One portion was not treated. Dimethylsulfoxide (Me2SO) in McCoy's medium was added to the untreated buffy coats and to the ficoll-hypaque treated buffy coats rapidly in 1 to 2 minutes or slowly over 15 to 20 minutes. The cell suspensions were frozen in polyolefin plastic bags at 2-3 C/ min by placing the plastic bag in a -80 C freezer, or at 1 C/min by use of a graded freezing apparatus. The percentage of viable mononuclear cells recovered between the untreated buffy coat and the ficoll-hypaquue treated buffy coat depleted of contaminting red blood cells and granulocytes regardless of the rate of addition of he Me2S0-McCoy's solution and the rate of freezing. There was a significant difference in the number of viable MNC's recovered when comparing untreated buffy coat and ficoll-hypaque treated buffy coat due to a 22% loss of MNC's during ficoll-hypaque treatment.
Document Details
- Document Type
- Technical Report
- Publication Date
- Oct 01, 1982
- Accession Number
- ADA131624
Entities
People
- A. D. Gray
- A. J. Melaragno
- Albert J. Roy
- J. Dittmer
- L. T. Lavigne Jr.
Organizations
- Boston University