›› 2009, Vol. 40 ›› Issue (1): 103-107.doi:

• 论著 • Previous Articles     Next Articles

Application of hydrodynamics-based transgent togene transfection in rat liver

  

  1. 1. College of Life Science,He′nan Normal University,He′nan Xinxiang 453007, China;2. Co.construction Key Laboratory for Cell Differentiation and Regulation, He′nan Xinxiang 453007, China;3. College of Life Science,Xinjiang University,Wulumuqi 830046,China
  • Received:2008-06-19 Revised:2008-07-09 Online:2009-01-06
  • Contact: XU Cun-shuan

Abstract: Objective To study the condition and method of hydrodynamics-based transgene when applied to gene transfection in rat liver. Methods Inject different dosages and concentrations of green fluorescent protein (GFP) plasmid (pEGFP-C1) at different speeds, then collect 4 rats’ liver leaves at different time points after injection for frozen section, and to observe and quantify the GFP expression with a fluorescence microscope at 488 nm excitation wavelength. Results With one-off injection(the optimal conditions), plasmid concentration is 30mg/L, injection speed is 2ml/s, injection volume is 9% of rat body weight. At 6 hours after injection, GFP-positive cells take on the highest proportion of above 20%; GFP begin to gradually reduce since 24 hours, until 72 hours almost no GFP-positive cells are seen in any liver leaf. Briefly(under the above optimal conditions), GFP expression in each liver leaf is described as follows: about 30% for pedicel leaf, about 20% for all of left, middle and right leaves,and about 10%for tail leaf.Conclusion Conclusion Hydrodynamics-based gene delivery procedure can be well applied to transgene in the rat liver, with the appropriate conditions of this method being 30mg/L plasmid concentration, 9%rat avoirdupois, 2ml/s injection speed. The most suita

Key words: Liver, Green fluorescent protein, Hydrodynamics-based transgene, Rat

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