Gene marking in adeno-associated virus vector infected periosteum derived cells for cartilage repair.
OBJECTIVE
To evaluate both the potential for transferring genes to periosteal cells using an adeno-associated virus (AAV) vector and the potential for gene expression after transplantation of those cells to a cartilage defect in vivo.


METHODS
Periosteum was obtained from the tibia of 6-week-old rabbits and enzymatically digested. The isolated periosteum derived cells were cultured and the subconfluence cells were infected with a recombinant AAV expressing the LacZ gene (AAV-LacZ). Collagen gel containing the LacZ transferred, periosteum derived cells was transplanted into a full thickness articular cartilage defect in 10 rabbits.


RESULTS
Infected cells still growing on the plate continued to express LacZ at least 12 weeks after AAV infection, with the highest percentage of LacZ positive cells reaching 74.4%. The LacZ positive cells were recognized at the transplant sites in 8 out of 10 knees.


CONCLUSIONS
Gene expression in periosteum derived cells was sustained in vitro for at least 12 weeks using the AAV vector, and for 2 weeks ex vivo after transplantation into a cartilage defect.
Kobayashi
Naomi
N
Department of Orthopaedic Surgery, Yokohama City University School of Medicine, Yokohama, Japan. naomik@ea.mbn.or.jp
Koshino
Tomihisa
T
Uesugi
Masaaki
M
Yokoo
Naoki
N
Xin
Ke-Qin
KQ
Okuda
Kenji
K
Mizukami
Hiroaki
H
Ozawa
Keiya
K
Saito
Tomoyuki
T
D000260:Adenoviruses, Human / Q000235:genetics*; D000818:Animals; D002358:Cartilage, Articular / Q000201:enzymology / Q000293:injuries / Q000473:pathology*; D017690:Cell Transplantation; D002478:Cells, Cultured; D005819:Genetic Markers; D005822:Genetic Vectors / Q000235:genetics*; D006801:Humans; D007596:Joints / Q000601:surgery; D007763:Lac Operon / Q000235:genetics; D010521:Periosteum / Q000166:cytology* / Q000201:enzymology / Q000821:virology; D011817:Rabbits; D014161:Transduction, Genetic; D014945:Wound Healing; D001616:beta-Galactosidase / Q000378:metabolism
D005819:Genetic Markers; D001616:beta-Galactosidase