Purification of cord blood lymphocytes

J Immunol Methods. 1996 Sep 9;195(1-2):43-8. doi: 10.1016/0022-1759(96)00089-0.

Abstract

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.

MeSH terms

  • Cell Separation / methods*
  • Fetal Blood / cytology*
  • Humans
  • Immunophenotyping
  • Leukocyte Common Antigens / analysis
  • Lymphocytes / cytology*
  • Lymphocytes / immunology

Substances

  • Leukocyte Common Antigens