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Gene transfer into intact fetal skeletal muscle grown in vitro
Author(s) -
Cardoso Sandra M. Pinto,
Mutch Philip,
Scotting Paul J.,
Wigmore Peter M.
Publication year - 2004
Publication title -
muscle and nerve
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.025
H-Index - 145
eISSN - 1097-4598
pISSN - 0148-639X
DOI - 10.1002/mus.20051
Subject(s) - electroporation , skeletal muscle , microbiology and biotechnology , biology , fetus , in vitro , gene , myocyte , gene expression , anatomy , genetics , pregnancy
Abstract 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. Muscle Nerve 30: 87–94, 2004