PMID: 7539467May 11, 1995Paper

Purification of human basophils and mast cells by multistep separation technique and mAb to CDw17 and CD117/c-kit

Journal of Immunological Methods
M WillheimP Valent

Abstract

Basophils and mast cells represent distinct cell lineages within the hemopoietic system. Based on the unique cell surface antigen profile of both cells, we have established methods which allow the reproducible purification to homogeneity (> 99%) of normal human basophil granulocytes from the peripheral blood and of mast cells from human dispersed tissues. Basophils (n = 9) were purified by current counterflow elutriation followed by depletion of monocytes with CD14 mAb conjugated to magnetic beads, and subsequent cell sorting for CD217+ cells. Basophil purity was 99.5 +/- 0.4% (range 98.7-99.9%). Mast cells were obtained from lung (n = 6), uterus (n = 1), mastocytosis bone marrow (n = 2), and human foreskin (n = 2). Mast cells were purified by collagenase digestion followed by current counterflow elutriation and sorting with CD117/c-kit mAb. Mast cell purity was 99.4 +/- 0.7% (range: 97.5-99.9%). Purified cells were more than 90% viable and were able to release histamine on induction with IgE plus anti-IgE. Furthermore, the PCR technique could be applied on pure cells and confirmed expression of high affinity IgE receptor (Fc epsilon R1) alpha chain mRNA. Thus, by combining isolation techniques including elutriation, magnetic c...Continue Reading

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Citations

Feb 14, 2004·Methods : a Companion to Methods in Enzymology·Peter ValentRudolf Valenta
Aug 6, 1998·Journal of Immunological Methods·P M ShahP O Schiøtz
Jul 20, 1999·Journal of Immunological Methods·K HaischH Haas
Jan 29, 2000·Journal of Immunological Methods·S TsangR R Schellenberg
Aug 19, 2010·Hybridoma·Ling YangJie Zhu
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Mar 21, 1997·The Journal of Biological Chemistry·C SillaberP Valent
May 20, 1999·American Journal of Hematology·F WimazalP Valent
Jul 31, 1998·Journal of Oral Pathology & Medicine : Official Publication of the International Association of Oral Pathologists and the American Academy of Oral Pathology·M FormanekJ Kornfehl
Jul 13, 2007·International Journal of Immunopathology and Pharmacology·A W HauswirthP Valent

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