Chest
Laboratory and Animal InvestigationsPleural Space as a Site of Ectopic Gene Delivery: Transfection of Pleural Mesothelial Cells With Systemic Distribution of Gene Product
Section snippets
Materials and Methods
The Vanderbilt University Institutional Animal Care and Use Committee approved the study protocol.
In Vitro Transfection of Mesothelial Cells
The PALP plasmid successfully transfected primary mesothelial cells in vitro (Fig 1). There was no detectable level of PALP prior to transfection. By 24 and 48 h after transfection, significant increases (16-fold and 17-fold, respectively) of PALP production were seen over the baseline levels, followed by a gradual decline at the later time points.
Pleural Lavage Fluid Levels of PALP in Treatment Rabbits vs Control Rabbits
PALP was recovered in significantly higher amounts in the pleural lavage fluid from rabbits in the treatment group than from those in the control
Discussion
The objective of the present short-term study was to assess the pleural space as a site for ectopic gene expression and for the systemic distribution of transgene products. We have successfully demonstrated the following: (1) the transfection of primary pleural mesothelial cells in vitro with plasmids, (2) the transfection of pleural mesothelial cells in vivo with the expression of the gene product in the pleural space, and (3) the subsequent systemic distribution of the gene product.
Ectopic
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This research was supported by the St. Thomas Foundation, Nashville, TN.