Journal article
Human lung mast cells: purification and characterization
The Journal of immunology (1950), v 129(6), pp 2662-2667
Dec 1982
PMID: 6183340
Featured in Collection : UN Sustainable Development Goals @ Drexel
Abstract
Detailed studies of the biochemistry and pharmacology of mast cell-mediated inflammatory disorders have been hampered by the inability to purify human mast cells. We now report techniques to purify human lung mast cells to apparent homogeneity. The major purification steps are: 1) dispersion of lung fragments into a single-cell suspension with enzyme combinations (pronase-chymopapain, collagenase-elastase); 2) partial purification by countercurrent centrifugation elutriation (CCE); and 3) affinity column chromatography. Enzymatic dispersion yielded suspensions with congruent to 10(6) mast cells per gram of lung parenchyma in purities of 1.2 to 9.7%. Dispersed mast cells responded comparably to those in parent lung fragments to challenge with anti-human IgG and pharmacologic agonists. Elutriation of lung cell suspensions yielded mast cell-enriched fractions with purities up to 70%. High purity mast cell fractions were combined, passively sensitized with purified human penicillin (BPO)-specific IgE, and purified by a BPO-affinity column chromatography procedure. Post elutriation mast cell purities of 29 +/- 3.5% were increased to 84 +/- 3% (range 65 to 98%) by the affinity column. Short-term (24 hr) culture of column-purified mast cells allowed adherence of non-mast cell contaminants to tissue culture plates, further increasing purity (up to 100%). Purified mast cells were intact and functional as assessed by dye exclusion, survival in short-term culture, IgE-mediated histamine release, and modulation of release by the pharmacologic agonists adenosine, IBMX, prostaglandin E2, and fenoterol.
Metrics
8 Record Views
Details
- Title
- Human lung mast cells: purification and characterization
- Creators
- E S SchulmanD W MacGlashan, JrS P PetersR P SchleimerH H NewballL M Lichtenstein
- Publication Details
- The Journal of immunology (1950), v 129(6), pp 2662-2667
- Publisher
- United States
- Grant note
- HL 07159 / NHLBI NIH HHS 1T32 AI-07147 / NIAID NIH HHS
- Resource Type
- Journal article
- Language
- English
- Academic Unit
- Pulmonary, Critical Care, and Sleep (Medicine)
- Web of Science ID
- WOS:A1982PR36900063
- Scopus ID
- 2-s2.0-0020422258
- Other Identifier
- 991014878165004721
UN Sustainable Development Goals (SDGs)
This publication has contributed to the advancement of the following goals:
InCites Highlights
Data related to this publication, from InCites Benchmarking & Analytics tool:
- Web of Science research areas
- Immunology