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In vitro and in vivo validation of human and goat chondrocyte labeling by green fluorescent protein lentivirus transduction

Miot, Sylvie and Gianni-Barrera, Roberto and Pelttari, Karoliina and Acharya, Chitrangada and Mainil-Varlet, Pierre and Juelke, Henriette and Jaquiery, Claude and Candrian, Christian and Barbero, Andrea and Martin, Ivan. (2010) In vitro and in vivo validation of human and goat chondrocyte labeling by green fluorescent protein lentivirus transduction. Tissue engineering. Part C, Methods, Vol. 16, H. 1. pp. 11-21.

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Official URL: http://edoc.unibas.ch/dok/A6004999

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Abstract

We investigated whether human articular chondrocytes can be labeled efficiently and for long-term with a green fluorescent protein (GFP) lentivirus and whether the viral transduction would influence cell proliferation and tissue-forming capacity. The method was then applied to track goat articular chondrocytes after autologous implantation in cartilage defects. Expression of GFP in transduced chondrocytes was detected cytofluorimetrically and immunohistochemically. Chondrogenic capacity of chondrocytes was assessed by Safranin-O staining, immunostaining for type II collagen, and glycosaminoglycan content. Human articular chondrocytes were efficiently transduced with GFP lentivirus (73.4 +/- 0.5% at passage 1) and maintained the expression of GFP up to 22 weeks of in vitro culture after transduction. Upon implantation in nude mice, 12 weeks after transduction, the percentage of labeled cells (73.6 +/- 3.3%) was similar to the initial one. Importantly, viral transduction of chondrocytes did not affect the cell proliferation rate, chondrogenic differentiation, or tissue-forming capacity, either in vitro or in vivo. Goat articular chondrocytes were also efficiently transduced with GFP lentivirus (78.3 +/- 3.2%) and maintained the expression of GFP in the reparative tissue after orthotopic implantation. This study demonstrates the feasibility of efficient and relatively long-term labeling of human chondrocytes for co-culture on integration studies, and indicates the potential of this stable labeling technique for tracking animal chondrocytes for in cartilage repair studies.
Faculties and Departments:03 Faculty of Medicine > Departement Biomedizin > Department of Biomedicine, University Hospital Basel > Cell and Gene Therapy (Banfi)
03 Faculty of Medicine > Departement Biomedizin > Department of Biomedicine, University Hospital Basel > Tissue Engineering (Martin)
03 Faculty of Medicine > Bereich Operative Fächer (Klinik) > Kopfbereich > Kiefer- und Gesichtschirurgie (Zeilhofer)
03 Faculty of Medicine > Departement Klinische Forschung > Bereich Operative Fächer (Klinik) > Kopfbereich > Kiefer- und Gesichtschirurgie (Zeilhofer)
UniBasel Contributors:Martin, Ivan and Jaquiéry, Claude Armand and Barbero, Andrea
Item Type:Article, refereed
Article Subtype:Research Article
Bibsysno:Link to catalogue
Publisher:Mary Ann Liebert
ISSN:1937-3384
Note:Publication type according to Uni Basel Research Database: Journal article. This is a copy of an article published in the Tissue engineering © 2007 [copyright Mary Ann Liebert, Inc.]; Tissue engineering is available online at: http://online.liebertpub.com.
Language:English
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Last Modified:31 Dec 2015 10:52
Deposited On:26 Apr 2013 07:01

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