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Combination of baculovirus-expressed BMP-2 and rotating-shaft bioreactor culture synergistically enhances cartilage formation.
- Source :
-
Gene therapy [Gene Ther] 2008 Feb; Vol. 15 (4), pp. 309-17. Date of Electronic Publication: 2007 Dec 06. - Publication Year :
- 2008
-
Abstract
- Baculovirus is an emerging gene delivery vector, thanks to a number of unique advantages. Herein, we genetically modified the rabbit articular chondrocytes with a recombinant baculovirus (Bac-CB) encoding bone morphogenetic protein-2 (BMP-2), which conferred high level BMP-2 expression and triggered the re-differentiation of dedifferentiated third passage (P3) chondrocytes in the monolayer culture. The transduced and mock-transduced P3 cells were seeded into porous scaffolds and cultured in either the dishes or the rotating-shaft bioreactor (RSB), a novel bioreactor imparting a dynamic, two-phase culture environment. Neither mock-transduced constructs in the RSB culture nor the Bac-CB-transduced constructs in the static culture grew into uniform cartilaginous tissues. Only the Bac-CB-transduced constructs cultured in the RSB for 3 weeks resulted in cartilaginous tissues with hyaline appearance, uniform cell distribution, cartilage-specific gene expression and considerably enhanced cartilage-specific extracellular matrix deposition, as determined by histological staining, reverse transcription-PCR analyses and biochemical assays. This is the first study demonstrating that combination of baculovirus-mediated growth factor expression and RSB culture synergistically enhanced in vitro creation of cartilaginous tissues from dedifferentiated chondrocytes. Since baculovirus transduction is generally considered safe, this approach represents a viable alternative to stimulate the formation of engineered cartilage in a more cost-effective way than the growth factor supplementation.
- Subjects :
- Animals
Bone Morphogenetic Protein 2
Cell Differentiation
Cell Line
Enzyme-Linked Immunosorbent Assay
Rabbits
Reverse Transcriptase Polymerase Chain Reaction
Tissue Engineering
Baculoviridae genetics
Bioreactors
Bone Morphogenetic Proteins genetics
Cartilage growth & development
Genetic Vectors
Transforming Growth Factor beta genetics
Subjects
Details
- Language :
- English
- ISSN :
- 1476-5462
- Volume :
- 15
- Issue :
- 4
- Database :
- MEDLINE
- Journal :
- Gene therapy
- Publication Type :
- Academic Journal
- Accession number :
- 18059504
- Full Text :
- https://doi.org/10.1038/sj.gt.3303087