Control of fibroblast contamination in primary rat skeletal muscle cell cultures: Effects of an epidermal growth factor linked cytotoxin
AuthorPierce, Paul Randall, 1951-
AdvisorAllen, Ronald E.
MetadataShow full item record
PublisherThe University of Arizona.
RightsCopyright © is held by the author. Digital access to this material is made possible by the University Libraries, University of Arizona. Further transmission, reproduction or presentation (such as public display or performance) of protected items is prohibited except with permission of the author.
AbstractThe in vitro study of muscle cell growth is hampered by the presence of non-muscle cells, particularly fibroblasts. The heterobifunctional cross-linking agent, N-succinimidyl 3-(2-pyridyldithio) propionate (SPDP) has been used to create a novel "toxic growth factor" to address the problem. Epidermal growth factor (EGF), which stimulates fibroblast but not satellite cell proliferation, was conjugated via SPDP to a potent ribosome inhibitor, pokeweed antiviral protein (PAP). By preferentially binding to fibroblasts, it was hoped that EGF-PAP could cytotoxically eliminate fibroblasts from primary cultures of rat skeletal muscle satellite cells. While EGF-PAP did prove to be a fibroblast cytotoxin, it could not completely eliminate them from cell cultures. Low dose-time exposures improved the ratio of multinucleated cells to mononucleated cells (percent fusion) by up to 66% over controls, but increased concentrations, or durations of EGF-PAP treatment, proved detrimental to satellite cell growth and/or differentiation.
Degree ProgramGraduate College