Gene delivery to human chondrocytes by an adeno associated virus vector.
OBJECTIVE
To investigate the efficiency of gene transduction to human chondrocytes using an adeno associated virus (AAV) vector.


METHODS
We transduced green fluorescent protein (GFP) gene using AAV vector to primary human chondrocytes as well as human cartilage organ cultures, in which chondrocytes are surrounded by extracellular matrix. Expression of GFP gene was analyzed at various time points after transduction by fluorescence microscopy and immunohistochemistry.


RESULTS
In primary chondrocytes, the percentages of GFP positive cells were 15.9% or 16.0% on Day 1 and 95.0% or 93.7% on Day 7 after gene transduction. In cartilage organ cultures, gene delivery was observed in cells located not only in the superficial layer but also in the deep layer within the cartilage tissue. Up to 45.3+/-7.4% or 46.0+/-3.9% of chondrocytes expressed GFP for at least 28 days.


CONCLUSIONS
AAV vector could be useful for direct gene delivery to chondrocytes in situ.
Arai
Y
Y
Department of Orthopaedic Surgery, Research Institute for Neurological Diseases and Geriatrics, Kyoto Prefectural University of Medicine, Japan.
Kubo
T
T
Fushiki
S
S
Mazda
O
O
Nakai
H
H
Iwaki
Y
Y
Imanishi
J
J
Hirasawa
Y
Y
D000256:Adenoviridae / Q000235:genetics*; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D002356:Cartilage / Q000166:cytology / Q000502:physiology; D002478:Cells, Cultured; D019902:Chondrocytes / Q000166:cytology / Q000502:physiology*; D005260:Female; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D015316:Genetic Therapy; D049452:Green Fluorescent Proteins; D006801:Humans; D007202:Indicators and Reagents / Q000378:metabolism; D008164:Luminescent Proteins / Q000235:genetics; D008297:Male; D009924:Organ Culture Techniques; D010003:Osteoarthritis / Q000628:therapy
D007202:Indicators and Reagents; D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins