Collection, Separation, Cryopreservation and Characterization of Peripheral Blood and Bone Marrow Stem Cells and Their Use in Treating Lethally Irradiated Dogs.
Annual rept. 1 Jun 82-31 May 83,
NAVAL BLOOD RESEARCH LAB BOSTON MASS
Pagination or Media Count:
Peripheral blood mononuclear cells MNC were obtained from buffy coats collected during 26 platelet apheresis procedures in the dog using the Haemonetics 30 Blood Processor. The number of mononuclear cells and platelets were determined for each procedure. After separation of the platelet rich plasma, dimethylsulfoxide Me2SO in McCoys medium was added to the residual 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 per minute by placing the plastic bag in a -80 C freezer or at 1 C per minute by use of a graded freezing apparatus. The percentage of viable MNCs recovered was determined after thawing and washing by measurement of uptake of fluorescein deacetate and thidium bromide. The average harvest of platelets from a standardized four pass procedure was 1.26 x 10 to the 11th power cells and of MNCs 1,340,000,000 cells. Viable recovery of frozen, thawed and washed MNCs was similar when frozen at 1 Cminute with slow addition of the cryophylactic agent to those frozen at 2-3 Cminute with rapid addition.
- Medicine and Medical Research