Gene transfer into intact fetal skeletal muscle grown in vitro.
The development of an organ culture system for growing prenatal intercostal muscle in vitro and its use to study gene function is described. Fetal skeletal muscle is relatively inaccessible during the key stages of its development, and this method enables DNA transfections and other manipulations to be carried out. The system allows cell proliferation and differentiation to continue and also maintains the morphology and fiber types of developing muscle. Gene transfer into cultured embryonic intercostal muscle was achieved by square-pulse electroporation of intact pieces of tissue. Expression of a marker gene (GFP) was found within 5 h and maintained for 2 days in muscle fibers and cells. The technique should enable the function of genes implicated in muscle development and disease to be studied at stages when access is difficult and in a controlled environment.
Cardoso
Sandra M Pinto
SM
School of Biomedical Sciences, University of Nottingham, Queen's Medical Centre, Nottingham, NG7 2UH, UK.
Mutch
Philip
P
Scotting
Paul J
PJ
Wigmore
Peter M
PM
D000818:Animals; D002454:Cell Differentiation; D002455:Cell Division; D018274:Electroporation / Q000379:methods; D005260:Female; D005333:Fetus; D018485:Muscle Fibers, Skeletal / Q000166:cytology / Q000502:physiology; D018482:Muscle, Skeletal / Q000166:cytology / Q000502:physiology*; D009924:Organ Culture Techniques / Q000379:methods*; D011247:Pregnancy; D051381:Rats; D017208:Rats, Wistar; D014162:Transfection / Q000379:methods*