Transfection of epithelial cells is enhanced by combined treatment with mannitol and polyethyleneglycol

The Journal of Gene Medicine
M DüchlerE Wagner

Abstract

Gene transfer efficiency drops significantly when polarized mammary epithelial cells are transfected instead of actively growing cells. However, fully differentiated cells are the targets for gene transfer in many in vivo applications. Therefore, a simple and effective method for the transfection of polarized mammary epithelial cells in confluent monolayers was developed. Reporter gene plasmids were complexed with polyethylenimine with an average molecular weight of 25 kDa (PEI 25), or other agents, to transfect confluent monolayers of ovine mammary epithelial cells (OMEC II) or human carcinoma cells (CaCo-2) in vitro. The improved technique included pretreatment of the cells with a hyperosmotic mannitol solution (7%) which caused a loosening of the tight contacts between the cells. Alternatively, the mannitol shock could be replaced by a short treatment with trypsin or EDTA. In addition to the pretreatment, 12.5% polyethyleneglycol with an average molecular weight of 8000 kDa (PEG 8000) was included in the transfection mixture containing the DNA complexes. The combined application of mannitol and PEG resulted in a very reliable 5- to 30-fold increase in reporter gene expression in OMEC II and CaCo-2 cells, but not K562 cells (...Continue Reading

References

Sep 15, 1976·The Biochemical Journal·B MaggioJ A Lucy
Jul 1, 1992·Proceedings of the National Academy of Sciences of the United States of America·E WagnerM L Birnstiel
Aug 1, 1995·Proceedings of the National Academy of Sciences of the United States of America·O BoussifJ P Behr
Feb 1, 1993·Analytical Biochemistry·F W Kluxen, H Lübbert
Feb 24, 1997·Biochemical and Biophysical Research Communications·S Li, J Thacker
Jan 1, 1997·Critical Reviews in Therapeutic Drug Carrier Systems·R J Lee, L Huang
Jan 1, 1997·Critical Reviews in Therapeutic Drug Carrier Systems·B Illel
Apr 29, 1998·The Journal of Biological Chemistry·H PollardD Escande
Aug 6, 1998·Biology of the Cell·M DüchlerK Schellander
Feb 6, 1999·Biotechnology Progress·S T AndreadisM L Yarmush
Apr 13, 1999·The American Journal of Physiology·T A Reinhardt, R L Horst
May 29, 1999·Brazilian Journal of Medical and Biological Research = Revista Brasileira De Pesquisas Médicas E Biológicas·C Ropert
Aug 3, 1999·Biological Chemistry·H Büeler
Aug 28, 1999·Molecular Biotechnology·N S Templeton, D D Lasic
Sep 24, 1999·Advances in Genetics·R I MahatoA Rolland

❮ Previous
Next ❯

Citations

Jul 12, 2002·The Journal of Gene Medicine·Markus DüchlerMathias Müller
May 25, 2013·European Journal of Pharmaceutics and Biopharmaceutics : Official Journal of Arbeitsgemeinschaft Für Pharmazeutische Verfahrenstechnik E.V·Wanling LiangJenny K W Lam
Jun 30, 2006·The Journal of Gene Medicine·H R MellorR Callaghan
Dec 17, 2014·Molecular Pharmaceutics·Santosh K MisraDipanjan Pan

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.

Related Papers

Therapeutic Delivery
Jamie L BetkerThomas J Anchordoquy
Journal of Aerosol Medicine and Pulmonary Drug Delivery
Shuhua BaiFakhrul Ahsan
Journal of Biochemical and Biophysical Methods
Bibiana D RiquelmeJean-François Stoltz
© 2021 Meta ULC. All rights reserved