Somatic gene transfer into the lactating ovine mammary gland.
BACKGROUND
Somatic gene therapy requires safe and efficient techniques for the gene transfer procedure. The ovine mammary gland is described as a model system for the evaluation of somatic gene transfer methods.


METHODS
Different gene delivery formulations were retrogradely injected into the mammary gland of lactating sheep. The efficiency of the gene transfer was subsequently measured by the detection of the secreted transgene products in the milk. To counteract the milk flow in the lactating gland caused by the permanent milk production, a newly developed pretreatment of the mammary gland with hyperosmotic solutions was applied. In addition, in vivo electroporation of DNA into the mammary gland is described.


RESULTS
Gene transfer using naked DNA or simple complexes of DNA with polycations did not result in traceable amounts of reporter gene products. However, utilizing the complex cationic lipid DOSPER, a peak expression of about 400 ng/ml was observed 6 days after transfection. Maximum expression rates of more than 1 microg/ml were obtained by combining hyperosmotic pretreatment and receptor-mediated gene transfer. For the in vivo electroporation, the proof of principle for this technique in the mammary gland is reported.


CONCLUSIONS
The ovine mammary gland turned out to be a very well suited as a model system for evaluation and optimization of various gene transfer protocols.
Institute of Biotechnology in Animal Production, University of Veterinary Medicine, IFA Tulln, Konrad Lorenzstrasse 20, A-3430 Tulln, Austria. markus.duechler@univie.ac.at
Markus
M
Düchler
Margret
M
Pengg
Susanne
S
Schüller
Franz
F
Pfneisl
Christoph
C
Bugingo
Gottfried
G
Brem
Ernst
E
Wagner
Karl
K
Schellander
Mathias
M
Müller
D000818:Animals; D018274:Electroporation; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D007774:Lactation; D008321:Mammary Glands, Animal / Q000033:anatomy & histology / Q000378:metabolism*; D009994:Osmolar Concentration; D012756:Sheep; D012996:Solutions; D014162:Transfection
D012996:Solutions