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Titolo:
High-efficiency non-viral transfection of primary chondrocytes and perichondrial cells for ex-vivo gene therapy to repair articular cartilage defects
Autore:
Goomer, RS; Deftos, LJ; Terkeltaub, R; Maris, T; Lee, MC; Harwood, FL; Amiel, D;
Indirizzi:
Univ Calif San Diego, Sch Med, Dept Orthoped, La Jolla, CA 92093 USA Univ Calif San Diego La Jolla CA USA 92093 thoped, La Jolla, CA 92093 USA Univ Calif San Diego, Sch Med, Dept Med, La Jolla, CA 92093 USA Univ CalifSan Diego La Jolla CA USA 92093 pt Med, La Jolla, CA 92093 USA Vet Adm Med Ctr, La Jolla, CA USA Vet Adm Med Ctr La Jolla CA USAVet Adm Med Ctr, La Jolla, CA USA Seoul Natl Univ, Coll Med, Dept Orthoped Surg, Seoul, South Korea Seoul Natl Univ Seoul South Korea ept Orthoped Surg, Seoul, South Korea
Titolo Testata:
OSTEOARTHRITIS AND CARTILAGE
fascicolo: 3, volume: 9, anno: 2001,
pagine: 248 - 256
SICI:
1063-4584(200104)9:3<248:HNTOPC>2.0.ZU;2-7
Fonte:
ISI
Lingua:
ENG
Soggetto:
HORMONE-RELATED PEPTIDE; LIPOSOME-MEDIATED TRANSFECTION; ENDOCHONDRAL BONE-FORMATION; GROWTH-FACTOR-BETA; IN-VITRO; IMMUNE-RESPONSES; CYSTIC-FIBROSIS; COLLAGEN GENE; EXPRESSION; ADENOVIRUS;
Keywords:
gene therapy; tissue engineering; articular cartilage repair; transfection; primary perichondrial cells; primary chondrocytes; PTHrP; TGF-beta 1;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Clinical Medicine
Life Sciences
Citazioni:
60
Recensione:
Indirizzi per estratti:
Indirizzo: Goomer, RS Univ Calif San Diego, Sch Med, Dept Orthoped, 9500 Gilman Dr, La Jolla, CA92093 USA Univ Calif San Diego 9500 Gilman Dr La Jolla CA USA 92093 3 USA
Citazione:
R.S. Goomer et al., "High-efficiency non-viral transfection of primary chondrocytes and perichondrial cells for ex-vivo gene therapy to repair articular cartilage defects", OSTEO CART, 9(3), 2001, pp. 248-256

Abstract

Background: Primary perichondrial cells and chondrocytes have been used torepair articular cartilage defects in tissue engineering studies involvingvarious animal models. Transfection of these cells with a gene that induces chondrocytic phenotype may form an ideal method to affect tissue engineering of articular cartilage. Design: A protocol for high-efficiency transfection of primary perichondrial and cartilage cells was optimized. Plasmids carrying the marker beta -galactosidase (beta -gal), PTHrP and TGF-beta1 genes driven by a strong mammalian promoter were transfected into primary perichondrial cells and chondrocytes. A three-step method was used to achieve high efficiency of transfection: (1) permeabilization of primary cells using a mild detergent, (2) association of plasmid DNAs with a polycationic (poly-l-lysine) core covalentlylinked to a receptor ligand (transferrin), (3) introduction of cationic liposomes to form the quaternary complex. For in-vivo assessment, polylactic acid (PLA) scaffolds seeded with beta -gal transfected perichondrial cells were implanted into experimentally created osteochondral defects in rabbit knees for 1 week. Results: The efficiency of transfection was determined to be over 70% in vitro. The transformed cells continued to express beta -gal, in vivo for theentire test period of 7 days. Furthermore, primary perichondrial cells transfected with TGF-beta1 and PTHrP over-expressed their cognate gene products. Conclusion: The ability to transfect autologous primary perichondrial cells and chondrocytes with high efficiency using a non-viral system may form afirst step towards tissue engineering with these transformed cells to repair articular cartilage defects. (C) 2001 OsteoAtthritis Research Society International.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 05/07/20 alle ore 22:43:00