Adenovirus-mediated gene transfection in the isolated lymphatic vessels

Methods Mol Biol. 2012;843:199-204. doi: 10.1007/978-1-61779-523-7_19.

Abstract

The authors describe technical details of experimental protocol of gene transfection in isolated rat mesenteric lymphatic vessels (MLVs). Authors also refer to the recent publication in Microcirculation, which provides wide set of experimental evidences obtained from confocal microscopic imaging and isolated vessels functional tests, which confirmed a successful achievement of the following goals. (1) Optimization of the experimental conditions to maintain the isolated "normal" rat mesenteric vessels in culture for sufficiently long periods of time to permit effective knockdown or overexpression of selected proteins/genes. (2) Development of the effective transfection protocols for lymphatic muscle and/or endothelial cells in intact isolated rat MLVs without nonspecific impairment on lymphatic contractile function due to the transfection protocol per se.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Adenoviridae / genetics*
  • Animals
  • Dissection
  • Genes, Reporter / genetics
  • Green Fluorescent Proteins / genetics
  • Lymphatic Vessels / metabolism*
  • Lymphatic Vessels / surgery
  • Male
  • Rats
  • Rats, Sprague-Dawley
  • Transfection / methods*

Substances

  • Green Fluorescent Proteins