| Enrichment of subpopulations of respiratory epithelial cells using flow cytometry. | |
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MedLine Citation:
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PMID: 1846747 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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The application of flow cytometry to enrich airway epithelial cell subpopulations is described. A complementary epithelial cell preparative technique is also outlined. The ability of the airway epithelium to protect the lung from environmental insults results from a complex interaction among the different cells that form its matrix. The separation of the different epithelial cell types is an essential step in the studies of mechanisms of the controlling factors of cell repair, cell differentiation, and neoplastic transformation. Epithelial cells of the New Zealand white rabbit trachea are prepared using enzymatic digestion and microdissection. Small sections of tracheal wall are dissected into pieces approximately 10 mm2. The mucosa is dissected and placed in 0.15% hyaluronidase for 40 min at 22 degrees C. Mucus is removed, and the mucosa is then placed in 0.1% pronase at 37 degrees C for 30 min. With careful dissection, the epithelium can be dissected from the mucosa in 10-mm2 sheets. Sheets of epithelial cells are placed in 6 ml of an enzymatic solution containing collagenase, 0.2% bovine serum albumin, 0.04% soya bean trypsin inhibitor, 0.06 ml of 1 M Hepes buffer for 3 h at 37 degrees C. The cells are gently pipetted during the 3-h period, yielding a suspension of viable cells. Subpopulations of these different cell types are enriched using an Orthocytofluorograph 50111. A krypton ion laser was used for excitation of cells at 488 nm. Forward-angle and 90 degrees scatter were gated on the histogram. The purification of the ciliated, basal, and secretory cells was 90%, 97%, and 94%, respectively.(ABSTRACT TRUNCATED AT 250 WORDS) |
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Authors:
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M L Aitken; M Villalon; P Verdugo; M Nameroff |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S. |
Journal Detail:
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Title: American journal of respiratory cell and molecular biology Volume: 4 ISSN: 1044-1549 ISO Abbreviation: Am. J. Respir. Cell Mol. Biol. Publication Date: 1991 Feb |
Date Detail:
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Created Date: 1991-03-08 Completed Date: 1991-03-08 Revised Date: 2007-11-14 |
Medline Journal Info:
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Nlm Unique ID: 8917225 Medline TA: Am J Respir Cell Mol Biol Country: UNITED STATES |
Other Details:
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Languages: eng Pagination: 174-8 Citation Subset: IM |
Affiliation:
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Department of Medicine, University of Washington, Seattle 98195. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Animals Cell Separation / methods* Cell Survival Electrophoresis, Polyacrylamide Gel Epithelial Cells Epithelium / metabolism, ultrastructure Female Flow Cytometry / methods* Hyaluronoglucosaminidase / metabolism Microbial Collagenase / metabolism Microscopy, Electron Pronase / metabolism Rabbits Trachea / cytology*, metabolism, ultrastructure Trypsin Inhibitors / metabolism |
| Grant Support | |
ID/Acronym/Agency:
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HL-01920/HL/NHLBI NIH HHS |
| Chemical | |
Reg. No./Substance:
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0/Trypsin Inhibitors; EC 3.2.1.35/Hyaluronoglucosaminidase; EC 3.4.24.-/Pronase; EC 3.4.24.3/Microbial Collagenase |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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