Method for umbilical cord blood-derived basophils by FCM

Hybridoma (Larchmt). 2010 Aug;29(4):367-70. doi: 10.1089/hyb.2010.0004.

Abstract

We have devised a method for the purification of human basophils from umbilical cord blood by FCM. Umbilical cord blood was collected from six healthy, full-term deliveries. After separation of red blood cells, mononuclear cells were isolated by Ficoll-Hypaque. Six samples followed by positive selection using flowcytometry (FCM) for CD203c(+)CD45(int+) cells. Purity and recovery of cells were measured. Purity and recovery of basophils by FCM with CD203c(+)CD45(int+) markers were 95.02 +/- 2.94% and 61.42 +/- 5.95%. Cell sorting for CD203c(+)CD45(int+) cells by FCM is an improved method for obtaining pure umbilical cord blood-derived basophils. We established cord blood-derived basophil purification technique as a source from which active basophils can be isolated for immunochemical characterization.

MeSH terms

  • Basophils / cytology*
  • Basophils / physiology
  • Cell Separation
  • Fetal Blood / cytology*
  • Ficoll / chemistry
  • Flow Cytometry*
  • Humans
  • Leukocyte Common Antigens / immunology
  • Leukocyte Common Antigens / metabolism
  • Leukocytes / cytology
  • Leukocytes / immunology
  • Phosphoric Diester Hydrolases / immunology
  • Phosphoric Diester Hydrolases / metabolism
  • Pyrophosphatases / immunology
  • Pyrophosphatases / metabolism

Substances

  • ENPP3 protein, human
  • Ficoll
  • Leukocyte Common Antigens
  • Phosphoric Diester Hydrolases
  • Pyrophosphatases