NDLI: Efficient and rapid isolation and purification of mouse alveolar type II epithelial cells.

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Efficient and rapid isolation and purification of mouse alveolar type II epithelial cells.

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Efficient and rapid isolation and purification of mouse alveolar type II epithelial cells.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Messier, Elise M. Mason, Robert J. Kosmider, Beata
Description	Country affiliation: United States Author Affiliation: Messier EM ( Department of Medicine, National Jewish Health, Denver, Colorado, USA.)
Abstract	The alveolar surface is covered by an epithelium composed of 2 main cell types: type I and type II cells. Alveolar type II (ATII) cells have a distinct morphology with apical microvilli and characteristic lamellar bodies, which are the intracellular storage form of pulmonary surfactant. ATII cells play an important role in innate immunity and produce and secrete pulmonary surfactant. They proliferate to restore the epithelium after damage to the more sensitive type I cells. We developed an efficient and rapid method to isolate and purify ATII cells from mice. Alveolar epithelial cells were dissociated in the murine lung with dispase and lung tissue was gently minced with a GentleMACS Dissociator. ATII cell purification was performed using negative depletion with CD45 MicroBeads and positive selection for the epithelial-cell adhesion molecule (Ep-CAM) by magnetic labeling with Streptavidin MicroBeads in MACS LS columns. The purity of these cells as measured by flow cytometry was up to 92.1% and 91.1% for co-staining with Ep-CAM and cytokeratin and co-staining with Ep-CAM and SP-A, respectively. The resulting ATII cell population has a high purity, viability, and yield. The phenotype of isolated and cultured ATII cells was confirmed by electron micrographs, expression of surfactant proteins (SP-A, proSP-B, mature SP-B, proSP-C, SP-D), and lysophosphatidylcholine acyltransferase (LPCAT) by western blotting and immunocytofluorescence. This protocol is based on surface antigens and our data demonstrated that murine ATII cells can be rapidly isolated, efficiently purified, and effectively cultured.
File Format	HTM / HTML
ISSN	01902148
Issue Number	7
Volume Number	38
e-ISSN	15210499
Journal	Experimental Lung Research
Language	English
Publisher	Taylor & Francis
Publisher Date	2012-09-01
Publisher Place	Great Britain (UK)
Access Restriction	Subscribed
Subject Keyword	Discipline Pulmonary Medicine Cell Separation Methods Epithelial Cells Metabolism Pulmonary Alveoli 1-acylglycerophosphocholine O-acyltransferase Biosynthesis Animals Antigens, Surface Analysis Cells, Cultured Cytology Female Male Mice Mice, Inbred C57bl Microspheres Pulmonary Surfactant-associated Proteins Journal Article Research Support, N.i.h., Extramural Research Support, Non-u.s. Gov't
Content Type	Text
Resource Type	Article
Subject	Pulmonary and Respiratory Medicine Molecular Biology Clinical Biochemistry

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