Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.
The cells are maintained in a serum free medium. If grown in media containing serum, the properties of the cells will change.
The cells exhibit different characteristic when grown with or without retinoic acid.
In the presence of retinoic acid (50 nM), the cells resemble low non-keratinized or squamous epithelium with densely packed colonies. In the absence of retinoic acid, the cells form a keratinized epithelium.
Long term cultures in the absence of retinoic acid form a stratified highly keratinized epithelium with large networks of epithelial filaments.
Volumes used in this protocol are for 75 cm2 flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.
Subcultivation Ratio: A subcultivation ratio of 1:10 to 1:20 is recommended
Medium Renewal: Every 2 to 3 days
Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 10 in Culture of Animal Cells, a manual of Basic Technique by R. Ian Freshney, 3rd edition, published by Alan R. Liss, N.Y., 1994.
Mather JP, Roberts PE. Method of isolating lung cell line. US Patent 5,364,785 dated Nov 15 1994
Roberts PE, et al. A novel epithelial cell from neonatal rat lung: isolation and differentiated phenotype. Am. J. Physiol. 259: L415-L425, 1990. PubMed: 2260675
Hay, R. J., Caputo, J. L., and Macy, M. L., Eds. (1992), ATCC Quality Control Methods for Cell Lines. 2nd edition, Published by ATCC.
Caputo, J. L., Biosafety procedures in cell culture. J. Tissue Culture Methods 11:223-227, 1988.
Fleming, D.O., Richardson, J. H., Tulis, J.J. and Vesley, D., (1995) Laboratory Safety: Principles and Practice. Second edition, ASM press, Washington, DC.
