期刊
LABORATORY INVESTIGATION
卷 84, 期 6, 页码 727-735出版社
NATURE PUBLISHING GROUP
DOI: 10.1038/labinvest.3700095
关键词
type I and type II pneumocytes; cell identification; immunomagnetic separation; hyperoxia
资金
- NHLBI NIH HHS [HL-52146, HL-071628] Funding Source: Medline
There are no ideal cell lines available for alveolar epithelial type I and II cells (AEC I and II) at the present time. The current methods for isolating AEC I and II give limited purities. Here, we reported improved and reproducible methods for the isolation of highly pure AEC I and II from rat lungs. AEC I and II were released from lung tissues using different concentrations of elastase digestion. Macrophages and leukocytes were removed by rat IgG 'panning' and anti-rat leukocyte common antigen antibodies. For AEC 11 isolation, polyclonal rabbit anti-T1alpha (an AEC I apical membrane protein) antibodies were used to remove AEC I contamination. For AEC I isolation, positive immunomagnetic selection by polyclonal anti-T1alpha antibodies was used. The purities of AEC I and II were 91 +/- 4 and 97 +/- 1 %, respectively. The yield per rat was similar to 2 x 10(6) for AEC I and similar to 33 x 10(6) for AEC II. The viabilities of these cell preparations were more than 96%. The protocol for AEC II isolation is also suitable to obtain pure AEC II (93-95%) from hyperoxia-injured and recovering lungs. The purified AEC I and II can be used for gene expression profiling and functional studies. It also offers an important tool to the field of lung biology.
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