Long-term gene expression using the lentiviral vector in rat chondrocytes.
The optimal approach to a long-term stable transgene expression in chondrocytes has not been established. Recently, lentiviral vectors have been used for transfection of some cultured cell lines. Our study tests the hypothesis that lentiviral vectors lead to longer gene expression in primary chondrocytes. We transfected lentiviral and adenoviral vectors carrying the green fluorescence protein gene to chondrocytes at different infection rates and cultured them in collagen Type I gel for up to 6 weeks. We also transplanted the cells of gel-suspended chondrocytes into the backs of nude mice. The mRNA expression of collagen Type II and aggrecan core protein was tested by real time polymerase chain reaction. The morphologic features and proliferation of chondrocytes were observed. Lentiviral vectors could transfect the green fluorescence protein gene to chondrocytes and the adenoviral vector, and there was no influence on the proliferation and phenotype of the chondrocytes. The percentage of lentiviral green fluorescence protein positive cells was much greater than the adenoviral green fluorescence protein at the end of 6 weeks. Stable green fluorescence protein expression was observed only in the lentivirus-transfected implants. The gene transfected by the lentiviral vector can be expressed efficiently for a long time and may be useful for gene transfer in cartilage defect repair.
Lu
Fei-Zhou
FZ
Laboratory of Transplantation Immunology, National Research Institute for Child Health and Development, 2-10-1 Okura, Setagaya-ku, Tokyo, Japan.
Kitazawa
Yusuke
Y
Hara
Yuko
Y
Jiang
Jian-Yuan
JY
Li
Xiao-Kang
XK
D000818:Animals; D002455:Cell Division; D002478:Cells, Cultured; D019902:Chondrocytes / Q000166:cytology / Q000502:physiology* / Q000637:transplantation; D024042:Collagen Type I / Q000494:pharmacology; D003470:Culture Media / Q000494:pharmacology; D015870:Gene Expression; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000235:genetics; D016086:Lentivirus / Q000235:genetics*; D008297:Male; D051379:Mice; D008819:Mice, Nude; D010641:Phenotype; D051381:Rats; D017207:Rats, Sprague-Dawley; D014162:Transfection / Q000379:methods*; D019076:Transgenes / Q000235:genetics*
D024042:Collagen Type I; D003470:Culture Media; D049452:Green Fluorescent Proteins