Chapter 9. Isolation and Culture of Pulmonary Alveolar Epithelial Type II Cells

  1. R. Ian Freshney and
  2. Mary G. Freshney
  1. Leland G. Dobbs and
  2. Robert F. Gonzalez

Published Online: 28 APR 2002

DOI: 10.1002/0471221201.ch9

Culture of Epithelial Cells, Second Edition

Culture of Epithelial Cells, Second Edition

How to Cite

Dobbs, L. G. and Gonzalez, R. F. (2002) Isolation and Culture of Pulmonary Alveolar Epithelial Type II Cells, in Culture of Epithelial Cells, Second Edition (eds R. I. Freshney and M. G. Freshney), John Wiley & Sons, Inc., New York, USA. doi: 10.1002/0471221201.ch9

Editor Information

  1. CRC Beatson Laboratories, Glasgow, Scotland

Author Information

  1. Suite 150, University of California Laurel Heights Campus, 3333 California Street, San Francisco, CA 94118

Publication History

  1. Published Online: 28 APR 2002
  2. Published Print: 12 APR 2002

Book Series:

  1. Culture of Specialized Cells

Book Series Editors:

  1. R. Ian Freshney

Series Editor Information

  1. CRC Beatson Laboratories, Glasgow, Scotland

ISBN Information

Print ISBN: 9780471401216

Online ISBN: 9780471221203

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Keywords:

  • pulmonary type II epithelium;
  • epithelial cell culture;
  • lung;
  • type II cells;
  • type I cells;
  • KGF;
  • lung fibroblasts;
  • coculture;
  • organotypic culture;
  • differentiation;
  • panning;
  • filtration;
  • Papanicolaou;
  • Matrigel

Summary

This chapter outlines some of the difficulties associated with the culture of Type II cells, including the loss of Type II differentiation and the possibility of reversible transdifferentiation into Type I cells. The role of fibroblast and matrix interaction, and KGF are discussed in this context, and the shortcomings of several continuous cell lines considered. Type II cells are isolated from rat lung after perfusion with elastase followed by disaggregation and filtration through cotton gauze and nylon mesh. Panning on IgG-coated plates is used to deplete the lymphocytes and macrophages. Sections are provided on troubleshooting potential problems in the isolation procedure. Type II cells are recognized by characteristic staining with the modified Papanicolaou stain. Differentiation is induced by culture on Matrigel with KGF and by coculture with lung fibroblasts.