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Gene transfer to adult human lung tissue ex vivo
Abstract
The potential of gene therapy for treatment of lung disease remains unrealised. Early model systems often resulted in promising efficiency of gene transfer, only to prove irreproducible in the clinic. While problems such as induction of host immune responses and duration of expression also need to be addressed, it is now widely believed that alternative, relevant models which more accurately reflect gene transfer efficiencies in human lungs are urgently required. We report here on a human lung slice culture system to assess gene transfer to adult lung epithelium. A lacZ-expressing adenovirus (AdCA35lacZ) was used as a reporter vector. A solution of AdCA35lacZ was instilled via bronchioles into resected lung tissue, a route analogous to clinical administration. Following a 1 h incubation, the tissue was inflated with a 0.4% agarose solution, instilled via the same bronchioles. Once solidified, 500 microm slices of the tissue were prepared and cultured for 4 days. beta-Galactosidase staining revealed lacZ transgene expression in bronchiolar and alveolar cells of the lung slices throughout the 4 days in culture. This system, which can also be used to study other viral and liposome vectors, could prove to be a useful alternative model for assessing gene delivery to adult human lung epithelium.
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LacZ transgene expression in PCLS was observed in bronchiolar and alveolar cells after 4 days in culture.
“LacZ transgene expression in PCLS was observed in bronchiolar and alveolar cells after 4 days in culture (McBride et al., 2000).”
From Pulmonary fibrosis: Emerging diagnostic and therapeutic strategies (2023) · cites this paper as McBride et al., 2000
It is possible to generate PCLS from mice via intra-pulmonary instillation of a lacZ-expressing adenovirus.
“it is possible to generate PCLS from mice via intra-pulmonary instillation of a lacZ-expressing adenovirus.”
From Pulmonary fibrosis: Emerging diagnostic and therapeutic strategies (2023) · cites this paper as McBride et al., 2000
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Metadata
- Authors
- McBride, S., Rannie, D., & Harrison, D. J.
- Affiliations
- University of Edinburgh (GB)
- ORCID
- Harrison, D J
- Venue
- Gene Therapy
- Published
- 2000-04-01
- Volume
- 7
- Issue
- 8
- Pages
- 675-678
- ISSN
- 0969-7128 (ISSN-L)
- 1476-5462
- OpenAlex
- W2328062178
- Cited by
- 13
- References
- 19
- Citations by year
- 2023: 1 · 2021: 1 · 2020: 1 · 2016: 1 · 2015: 2 · 2013: 1 · 2012: 1
- FWCI
- 0.19
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Gene Therapy (publishedVersion)
- PubMed (publishedVersion)
- Maynooth University ePrints and eTheses Archive (Maynooth University) (acceptedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Virus-based gene therapy research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Interference and Gene Delivery
- Health Sciences › Medicine › Pulmonary and Respiratory Medicine › Inhalation and Respiratory Drug Delivery
- Keywords
- lung epithelium, gene transfer, adenoviral vector, lacZ reporter, beta-galactosidase expression, bronchioles, alveolar cells, transgene expression, lung disease, ex vivo model
- MeSH terms
- Adenoviridae, Adult, Animals, Epithelium, Genetic Vectors, Humans, Lac Operon, Lung, Genetic Therapy, Gene Expression, Gene Transfer Techniques, Culture Techniques, Mice
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