Murine epithelial cells: isolation and culture.
We describe an air-liquid interface primary culture method for murine tracheal epithelial cells on semi-permeable membranes, forming polarized epithelia with a high transepithelial resistance, differentiation to ciliated and secretory cells, and physiologically appropriate expression of key genes and ion channels. We also describe the isolation of primary murine nasal epithelial cells for patch-clamp analysis, generating polarised cells with physiologically appropriate distribution and ion channel expression. These methods enable more physiologically relevant analysis of murine airway epithelial cells in vitro and ex vivo, better utilisation of transgenic mouse models of human pulmonary diseases, and have been approved by the European Working Group on CFTR expression.
B.C. Research Institute, Room 381, University of British Columbia, 950 West 28th Avenue, Vancouver, BC, Canada V5Z 4H4. ddavidso@interchange.ubc.ca
Donald J
DJ
Davidson
Michael A
MA
Gray
Fiona M
FM
Kilanowski
Robert
R
Tarran
Scott H
SH
Randell
David N
DN
Sheppard
Barry E
BE
Argent
Julia R
JR
Dorin
D000818:Animals; D002478:Cells, Cultured; D004847:Epithelial Cells; D016591:Histocytological Preparation Techniques / Q000379:methods*; D008567:Membranes, Artificial; D051379:Mice; D023421:Models, Animal; D018408:Patch-Clamp Techniques / Q000379:methods; D020545:Respiratory Mucosa / Q000473:pathology*; D013048:Specimen Handling / Q000379:methods; D014132:Trachea / Q000166:cytology / Q000473:pathology
D008567:Membranes, Artificial