Stability of fresh and preserved fetal and adult lymphocyte cell surface markers.
Delay between blood collection and immunophenotyping of peripheral blood (PB) and umbilical cord blood (UCB) lymphocytes occurs frequently. Holding media address this problem, but there are few reports of their limitations. We tested the ability of Cyto-Chex (CC) to preserve the ability of lymphocyte subpopulations to be immunophenotyped after time delays. Ten UCB and 10 PB specimens were kept up to 48 hr and then placed in CC for 1 week, with removal of aliquots for staining with CD3, CD4, CD8, CD19, and CD45 at intervals. There were no significant changes in the number of nonpreserved PB or UCB cells stained with any of the reagents after 48 hr. Aside from a small decrease in UCB B-cell numbers, there were no changes in UCB or PB lymphocyte numbers after placing them in CC for 1 week. However, PB and UCB CD19+ cells and CD8+ T cells lost capacity for bright staining after 1 week in CC. The results suggest that UCB and PB can be held for up to 48 hr before being placed in CC and then kept for up to 1 week in CC with no decrease in T-cell numbers and only a minor reduction of B-cell numbers, albeit with a marked reduction in staining intensity.
Schumacher
M J
MJ
Department of Pediatrics, The Steele Memorial Children's Research Center, University of Arizona Health Sciences Center, Tucson 85724, USA. schumach@u.arizona.edu
Burkhead
T
T
D000328:Adult; D015703:Antigens, CD / Q000032:analysis*; D018941:Antigens, CD19 / Q000032:analysis; D015415:Biomarkers / Q000032:analysis*; D001793:Blood Preservation; D017252:CD3 Complex / Q000032:analysis; D015704:CD4 Antigens / Q000032:analysis; D016827:CD8 Antigens / Q000032:analysis; D004355:Drug Stability; D005260:Female; D005312:Fetal Blood / Q000166:cytology*; D005333:Fetus; D005434:Flow Cytometry; D006801:Humans; D016130:Immunophenotyping; D017493:Leukocyte Common Antigens / Q000032:analysis; D008214:Lymphocytes / Q000276:immunology*; D011247:Pregnancy; D013997:Time Factors
D015703:Antigens, CD; D018941:Antigens, CD19; D015415:Biomarkers; D017252:CD3 Complex; D015704:CD4 Antigens; D016827:CD8 Antigens; D017493:Leukocyte Common Antigens