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The isolation of rat alveolar type II cells: a simplified approach using Percoll density centrifugation

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Abstract

Colloidal silica-polyvinylpyrrolidinone solution (Percoll) is used in this study to purify type II alveolar pneumocytes obtained from elastase disaggregation of rat lung tissue. The crude lung cell suspension (containing 31.4±2.1% type II cells) was applied to a Percoll gradient established by centrifugation of phosphate-buffered saline with the Percoll. Color-coded density marker beads permitted the determination of the density of each resulting cellular layer. Harvesting of the type II cell-enriched layer yielded a cell population 92±2% pure for type II cells. Identification of type II cells was made using a modified Papanicolaou stain, a negative myeloperoxidase stain, negative leukocyte cell surface markers, and ultrastructural examination. Purification of crude rat lung cell suspensions with Percoll appears to be a rapid and inexpensive means of obtaining a replenishable type II cell population.

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Authors and Affiliations

  1. Division of Thoracic Diseases and Internal Medicine, Mayo Clinic and Mayo Foundation, 55905, Rochester, Minnesota, USA

    David M. Skillrud & William J. Martin

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  1. David M. Skillrud
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  2. William J. Martin
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Supported in part by a Rappaport Clinical Investigator Award and National Institutes of Health Grant No. HL-28029

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Skillrud, D.M., Martin, W.J. The isolation of rat alveolar type II cells: a simplified approach using Percoll density centrifugation. Lung 162, 245–252 (1984). https://doi.org/10.1007/BF02715652

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