Reconstitution of tracheal grafts with a genetically modified epithelium.
A rational approach to the development of gene therapies for cystic fibrosis requires a better understanding of the cellular targets for gene transfer in the airway epithelium. We have used recombinant retroviruses to study the dynamics and lineage relationships of a regenerating rat tracheal epithelium. Primary cultures of tracheal epithelial cells were exposed to lacZ-transducing retroviruses and subsequently seeded into denuded trachea that were implanted into BALB/c (nu/nu) mice. The grafts developed a fully differentiated mucociliary epithelium containing large clones of lacZ-expressing cells with virtually all cell types represented within each clone. These data are most consistent with gene transfer into a putative progenitor cell that is capable of extensive self renewal and pleuripotent development. Vector-specific variation in transgene expression was noted in the various cell types.
Engelhardt
J F
JF
Department of Internal Medicine, Howard Hughes Medical Institute, University of Michigan Medical Center, Ann Arbor 48109.
Allen
E D
ED
Wilson
J M
JM
D016475:3T3 Cells; D000199:Actins / Q000235:genetics; D000818:Animals; D002478:Cells, Cultured; D002645:Chickens; D004847:Epithelial Cells; D004848:Epithelium / Q000201:enzymology / Q000648:ultrastructure; D005822:Genetic Vectors; D007150:Immunohistochemistry; D008297:Male; D051379:Mice; D008854:Microscopy, Electron; D011401:Promoter Regions, Genetic; D011817:Rabbits; D051381:Rats; D011916:Rats, Inbred F344; D011973:Receptors, LDL / Q000235:genetics; D012091:Repetitive Sequences, Nucleic Acid; D012190:Retroviridae / Q000235:genetics; D014132:Trachea / Q000166:cytology / Q000637:transplantation*; D014162:Transfection; D001616:beta-Galactosidase / Q000235:genetics* / Q000378:metabolism
D000199:Actins; D011973:Receptors, LDL; D001616:beta-Galactosidase