Efficient gene expression in megakaryocytic cell line using nucleofection

To clarify the mechanism of platelet production from megakaryocytes, expression of target proteins by gene transfection was examined using various gene delivery techniques. Transfection into hematopoietic cells, including megakaryocytes, by conventional gene delivery techniques such as electroporati...

Full description

Saved in:
Bibliographic Details
Published in:International journal of pharmaceutics 2007-06, Vol.338 (1), p.157-164
Main Authors: Isakari, Yoshimasa, Harada, Yasuo, Ishikawa, Dai, Matsumura-Takeda, Kuniko, Sogo, Shinji, Ishida, Tatsuhiro, Taki, Takao, Kiwada, Hiroshi
Format: Article
Language:English
Subjects:
Biological and medical sciences
Cell Line
Electroporation
General pharmacology
Genetic Vectors
Humans
Liposomes
Medical sciences
MEG-01
Megakaryocytes
Megakaryocytes - metabolism
Nucleofection
Pharmaceutical technology. Pharmaceutical industry
Pharmacology. Drug treatments
Receptors, Thrombopoietin - genetics
Sendai virus - genetics
Transfection
Transfection - methods
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:To clarify the mechanism of platelet production from megakaryocytes, expression of target proteins by gene transfection was examined using various gene delivery techniques. Transfection into hematopoietic cells, including megakaryocytes, by conventional gene delivery techniques such as electroporation and lipofection are known to be difficult. In this study, in addition to electroporation and lipofection, we tested other gene-transfer methods (nucleofection, transfection using inactivated virus envelope, and transferrin-linked cationic polymer) with the green fluorescent protein (GFP) gene into the human megakaryocytic cell line MEG-01. We found that nucleofection, which uses a combination of special electrical parameters and specific solutions, was the best, judging from the expression ratio of GFP-positive cells (approximately 70% of cells) and low toxicity. The efficiency of GFP expression was not related to the amount of pDNA delivered into the MEG-01 cells. To verify the utility of nucleofection, the thrombopoietin (TPO) receptor c-mpl was transfected into MEG-01 cells. Transfected cells showed a higher responsiveness to TPO than mock-transfected MEG-01 cells. We propose that nucleofection is a useful method for transfecting target genes to megakaryocytic cells when addressing the mechanism of platelet production.
ISSN:0378-5173
1873-3476
DOI:10.1016/j.ijpharm.2007.01.042