In vitro and in vivo tracing of fluorescently tagged cell types in lung

Background:

Lung development processes encompass various cellular processes including cellular interactions like alveolarisation, septa formation, differentiation and trans differentiation processes. Similar to this, lung diseases like pulmonary fibrosis, pulmonary hypertension are characterized by tissue remodelling processes. The aim of our project is to visualize these interaction processes by specific fluorescent labelling of different cell types by single plasmid containing fluorochromes under control of the respective cell type specific promoter.

Methods:

Cloning, cell culture, transfection, immunofluorescence, transgenic mice development

Results:

We were able to clone different plasmids. We finished first single fluorochrome plasmid with specific promoter as well as single plasmid with two flurochrome under two specific promoters. The construction of single plasmid to label three different cell types with three different fluorescent proteins is under progress. The single color plasmids could be successfully tested in cell culture whereas in vitro testing of other constructs is in progress.

Outlook:

Our further work will focus on the in vitro studies, validating the specific expression in different cell types and the creation of transgenic mice for in vivo analysis of tissue specific labeling which will facilitate cell specific sorting, in vivo imaging as well as developmental/disease specific cellular and molecular mechanism studies.