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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%.

journal_name

Scand J Immunol

journal_title

Scandinavian journal of immunology

authors

van Overveld FJ,Terpstra GK,Bruijnzeel PL,Raaijmakers JA,Kreukniet J

doi

10.1111/j.1365-3083.1988.tb02316.x

subject

Has Abstract

pub_date

1988-01-01 00:00:00

pages

1-6

issue

1

eissn

0300-9475

issn

1365-3083

journal_volume

27

pub_type

杂志文章

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