Transfection of neonatal rat cochlear cells in vitro with an adenovirus vector.
A recombinant adenovirus vector containing a beta-galactosidase reporter gene was used to transfect neonatal rat organ of Corti or spiral ganglion explants in vitro. Infection at appropriate titers (10(6)-10(7) pfu/ml) transduced virtually all cells in the cultures after 72 hr. However, spiral ganglion neurons and cells in the inner hair cell regions of the organ of Corti showed the highest levels of expression. Viral titers that produced high levels of beta-galactosidase expression did not appear to damage the cultures, and did not inhibit neurite outgrowth from spiral ganglion cells. However, higher titers (10(8)-10(9) pfu/ml) clearly diminished explant viability and inhibited neurite extension. The results demonstrate that cochlear cells can be transfected successfully with an adenovirus vector, at viral titers which do not induce obvious signs of cellular damage or dysfunction.
Dazert
S
S
Department of Surgery/Otolaryngology, UCSD School of Medicine and Veterans Administration Medical Center, La Jolla 92093-0666, USA.
Battaglia
A
A
Ryan
A F
AF
D000256:Adenoviridae / Q000235:genetics*; D000257:Adenoviridae Infections / Q000473:pathology / Q000821:virology; D000818:Animals; D000831:Animals, Newborn / Q000502:physiology*; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D003051:Cochlea / Q000166:cytology* / Q000502:physiology; D046508:Culture Techniques; D017930:Genes, Reporter; D005822:Genetic Vectors / Q000235:genetics*; D016501:Neurites / Q000502:physiology / Q000648:ultrastructure; D016900:Neurofilament Proteins / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D013136:Spiral Ganglion / Q000473:pathology / Q000821:virology; D014162:Transfection / Q000235:genetics*; D001616:beta-Galactosidase / Q000096:biosynthesis / Q000235:genetics
D016900:Neurofilament Proteins; D001616:beta-Galactosidase