title: Generation of Mouse Lung Epithelial Cells.
abstract: Althoughin vivomodels are excellent for assessing various facets of whole organism physiology, pathology, and overall response to treatments, evaluating basic cellular functions, and molecular events in mammalian model systems is challenging. It is therefore advantageous to perform these studies in a refined and less costly setting. One approach involves utilizing cells derived from the model under evaluation. The approach to generate such cells varies based on the cell of origin and often the genetics of the cell. Here we describe the steps involved in generating epithelial cells from the lungs ofKrasLSL-G12D/+; p53LSL-R172/+mice (Kasinski and Slack, 2012). These mice develop aggressive lung adenocarcinoma following cre-recombinase dependent removal of a stop cassette in the transgenes and subsequent expression ofKra-G12Dand p53R172. While this protocol may be useful for the generation of epithelial lines from other genetic backgrounds, it should be noted that theKras; p53cell line generated here is capable of proliferating in culture without any additional genetic manipulation that is often needed for less aggressive backgrounds.
authors.affiliation: Molecular, Cellular and Developmental Biology Department, Yale University, New Haven, USA.
authors.forename: Andrea L
authors.initials: AL
authors.lastname: Kasinski
authors.affiliation: Molecular, Cellular and Developmental Biology Department, Yale University, New Haven, USA.
authors.forename: Frank J
authors.initials: FJ
authors.lastname: Slack