In VitroandIn VivoValidation of Human and Goat Chondrocyte Labeling by Green Fluorescent Protein Lentivirus Transduction
| Autor: | Karoliina Pelttari, Henriette Juelke, Chitrangada Acharya, Christian Candrian, Roberto Gianni-Barrera, Sylvie Miot, Pierre Mainil-Varlet, Claude Jaquiery, Ivan Martin, Andrea Barbero |
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| Rok vydání: | 2010 |
| Předmět: |
Green Fluorescent Proteins
Biomedical Engineering Type II collagen Mice Nude Medicine (miscellaneous) Bioengineering Biology Chondrocyte Green fluorescent protein Mice Transduction (genetics) Chondrocytes In vivo medicine Animals Humans 610 Medicine & health Collagen Type II Cell Proliferation Glycosaminoglycans Goats Cartilage Lentivirus Reproducibility of Results Cell Differentiation Chondrogenesis Immunohistochemistry Molecular biology Coculture Techniques medicine.anatomical_structure Phenazines 570 Life sciences biology Immunostaining |
| Zdroj: | Tissue Engineering Part C: Methods. 16:11-21 |
| ISSN: | 1937-3392 1937-3384 |
| DOI: | 10.1089/ten.tec.2008.0698 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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