Culture of conducting airway epithelial cells in serum-free medium

Cynthia B. Robinson, Reen Wu

Research output: Contribution to journalArticle

39 Scopus citations

Abstract

Reproducible techniques for the isolation and culture of conducting airway epithelial cells from various species including human are described. Tissues recovered after necropsy or surgery are treated with 0.1% protease at 4°C overnight. The epithelial lining cells are liberated from adventitia by flushing with ice-cold minimal essential medium containing 10% fetal bovine serum. The resulting suspension of single cells and cell clusters is centrifuged and then plated on type 1 collagen gel substratum in a serum-free F12 medium supplemented with insulin, transferrin, epidermal growth factor, cholera toxin, hydrocortisone (or dexamethasone), bovine hypothalamus extract, and retinol. Increased calcium concentration in the medium to 1 to 3 m M augments in vitro mucous cell differentiation. Both a squamouslike and mucociliary differentiation of the cultured epithelium can be identified by biochemical, immunohistologic, and morphologic means. The polarity of epithelial cell cultures is promoted by use of biphasic culture methods, in which epithelial cells are fed basally and are in direct contact with the air phase.

Original languageEnglish (US)
Pages (from-to)95-102
Number of pages8
JournalJournal of Tissue Culture Methods
Volume13
Issue number2
DOIs
StatePublished - Jun 1991

Keywords

  • airways
  • biphasic culture
  • differentiation
  • epithelial cell
  • mucous cell
  • serum-free medium

ASJC Scopus subject areas

  • Pathology and Forensic Medicine
  • Anatomy

Fingerprint Dive into the research topics of 'Culture of conducting airway epithelial cells in serum-free medium'. Together they form a unique fingerprint.

  • Cite this