Purification of cord blood lymphocytes.
When cord blood is separated using standard methods based on Ficoll-Hypaque, the mononuclear fraction is contaminated with erythrocytes and also with nucleated cells that do not express the leucocyte marker CD45. The contamination with CD45-negative cells can exceed 50%, and will interfere with phenotypic, mRNA or functional analysis. A large proportion of these cells are erythrocyte precursors. The contaminating cells may be removed by lysis with hypotonic ammonium chloride; when the cells are required for studies which are adversely affected by ammonium chloride (such as antigen processing), high purity can be attained by two rounds of density separation.
Ridings
J
J
Child Health Research Institute, Adelaide Women's and Children's Hospital, Australia.
Weedon
H
H
Ioannou
C
C
Flego
L
L
Macardle
P J
PJ
Zola
H
H
D002469:Cell Separation / Q000379:methods*; D005312:Fetal Blood / Q000166:cytology*; D006801:Humans; D016130:Immunophenotyping; D017493:Leukocyte Common Antigens / Q000032:analysis; D008214:Lymphocytes / Q000166:cytology* / Q000276:immunology
D017493:Leukocyte Common Antigens