Effects of growth surface on differentiation of cultures of human tracheal epithelium

Jonathan Widdicombe, L. A. Sachs, W. E. Finkbeiner

Research output: Contribution to journalArticle

26 Scopus citations

Abstract

Optical measurements from epithelial cells grown on clear solid surfaces (e.g., coverslips, petri dishes) are often compared with other measurements (e.g., short-circuit current; Isc) obtained from cells grown on opaque porous surfaces (inserts). However, the relative levels of differentiation of cells grown under the two conditions are usually unknown. To address this issue, we grew primary cultures of human tracheal epithelium on solid surfaces or on porous inserts and compared their total levels of protein and deoxyribonucleic acid, electrical properties in Ussing chambers, and ultrastructure. To measure ion transport across cells grown on solid supports, cells were grown on inserts placed on parafilm. Later, separation of insert from parafilm allowed the cells' Isc to be measured in Ussing chambers. Four different media were used. Cells grown in one medium showed very low levels of differentiation on all growth supports. In the other media, growth on inserts markedly enhanced differentiation as compared with solid supports. Baseline Isc of cells grown on either clear or opaque inserts was at least 30 times greater than that of cells grown on solid supports, though Isc with clear inserts averaged ∼30% lower than that with opaque inserts. We conclude that though differentiation of cells may vary slightly depending on the insert used, cells on any type of insert are much better differentiated than cells grown on solid surfaces. Thus, it is both possible and desirable to make all functional measurements on cells grown on clear porous supports.

Original languageEnglish (US)
Pages (from-to)51-55
Number of pages5
JournalIn Vitro Cellular and Developmental Biology - Animal
Volume39
Issue number1-2
DOIs
StatePublished - Jan 2003

Keywords

  • Ion transport
  • Porous-bottomed inserts
  • Short-circuit current
  • Transepithelial electrical resistance

ASJC Scopus subject areas

  • Developmental Biology
  • Clinical Biochemistry
  • Cell Biology

Fingerprint Dive into the research topics of 'Effects of growth surface on differentiation of cultures of human tracheal epithelium'. Together they form a unique fingerprint.

  • Cite this