Developing cell therapy techniques for respiratory disease: intratracheal delivery of genetically engineered stem cells in a murine model of airway injury.
Interest has increased in the use of exogenous stem cells to optimize lung repair and serve as carriers of a therapeutic gene for genetic airway diseases such as cystic fibrosis. We investigated the survival and engraftment of exogenous stem cells after intratracheal injection, in a murine model of acute epithelial airway injury already used in gene therapy experiments on cystic fibrosis. Embryonic stem cells and mesenchymal stem cells were intratracheally injected 24 hr after 2% polidocanol administration, when epithelial airway injury was maximal. Stem cells were transfected with reporter genes immediately before administration. Reporter gene expression was analyzed in trachea-lungs and bronchoalveolar lavage, using nonfluorescence, quantitative, and sensitive methods. Enzyme-linked immunosorbent assay quantitative results showed that 0.4 to 5.5% of stem cells survived in the injured airway. Importantly, no stem cells survived in healthy airway or in the epithelial lining fluid. Using 5-bromo-4-chloro-3-indolyl-beta-d-galactopyranoside staining, transduced mesenchymal stem cells were detected in injured trachea and bronchi lumen. When the epithelium was spontaneously regenerated, the in vivo amount of engrafted mesenchymal stem cells from cell lines decreased dramatically. No stem cells from primary culture were located within the lungs at 7 days. This study demonstrated the feasibility of intratracheal cell delivery for airway diseases with acute epithelial injury.
Leblond
Anne-Laure
AL
INSERM, UMR915, Institut du Thorax, Nantes, France.
Naud
Patrice
P
Forest
Virginie
V
Gourden
Clothilde
C
Sagan
Christine
C
Romefort
Bénédicte
B
Mathieu
Eva
E
Delorme
Bruno
B
Collin
Christine
C
Pagès
Jean-Christophe
JC
Sensebé
Luc
L
Pitard
Bruno
B
Lemarchand
Patricia
P
D000704:Analysis of Variance; D000818:Animals; D018893:Bronchoalveolar Lavage; D064987:Cell- and Tissue-Based Therapy / Q000379:methods*; D017931:DNA Primers / Q000235:genetics; D053595:Embryonic Stem Cells / Q000637:transplantation*; D004797:Enzyme-Linked Immunosorbent Assay; D005697:Galactosides; D005818:Genetic Engineering / Q000379:methods; D005822:Genetic Vectors; D007211:Indoles; D008297:Male; D045164:Mesenchymal Stem Cell Transplantation / Q000379:methods*; D051379:Mice; D012140:Respiratory Tract Diseases / Q000628:therapy*; D014132:Trachea / Q000166:cytology*; D014162:Transfection; D001616:beta-Galactosidase
D017931:DNA Primers; D005697:Galactosides; D007211:Indoles; D001616:beta-Galactosidase; C044888:5-bromo-4-chloro-3-indolyl beta-galactoside