title: The isolation of human lung mast cells by affinity chromatography.
abstract: A method of isolation has been developed to purify mast cells from human lung tissue. The purification steps are: (1) dispersion of human lung tissue in single-cell suspensions by enzymatic digestion, (2) partial purification by counterflow centrifugal elutriation, (3) Percoll gradient centrifugation, and (4) enrichment of the mast cells by affinity chromatography using anti-human IgE-Sepharose. Enzymatic dispersion yielded 0.6 +/- 0.2 x 10(6) mast cells per gram wet tissue with purities of 3.3 +/- 1.0% (mean +/- SEM n = 3). Elutriation and gradient centrifugation yielded 0.36 +/- 0.05 x 10(6) mast cells per gram lung tissue in fractions with purities of 30.8 +/- 10.7%. Enriched mast cell fractions were combined, and disposed of contaminating cells by affinity chromatography, thereby yielding 0.25 +/- 0.03 x 10(6) mast cells per gram lung tissue, and improving the purity to 75.3 +/- 8.3%. The purified mast cells were intact and vitality exceeded 95%. In this way from 1 g wet lung tissue 0.25 +/- 0.03 x 10(6) mast cells may be isolated with a mean recovery of 41.7 +/- 2.4% and a mean purity of 75.3 +/- 8.3%.
authors.affiliation: Department of Pulmonary Disease, State University Hospital, Utrecht, The Netherlands.
authors.forename: F J
authors.initials: FJ
authors.lastname: van Overveld
authors.forename: G K
authors.initials: GK
authors.lastname: Terpstra
authors.forename: P L
authors.initials: PL
authors.lastname: Bruijnzeel
authors.forename: J A
authors.initials: JA
authors.lastname: Raaijmakers
authors.forename: J
authors.initials: J
authors.lastname: Kreukniet
mesh_terms: D002469:Cell Separation / Q000379:methods*; D002499:Centrifugation, Density Gradient; D002846:Chromatography, Affinity; D006801:Humans; D008168:Lung / Q000166:cytology*; D008407:Mast Cells