title: Gene marking in adeno-associated virus vector infected periosteum derived cells for cartilage repair.
abstract: 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.
authors.affiliation: Department of Orthopaedic Surgery, Yokohama City University School of Medicine, Yokohama, Japan. naomik@ea.mbn.or.jp
authors.forename: Naomi
authors.initials: N
authors.lastname: Kobayashi
authors.forename: Tomihisa
authors.initials: T
authors.lastname: Koshino
authors.forename: Masaaki
authors.initials: M
authors.lastname: Uesugi
authors.forename: Naoki
authors.initials: N
authors.lastname: Yokoo
authors.forename: Ke-Qin
authors.initials: KQ
authors.lastname: Xin
authors.forename: Kenji
authors.initials: K
authors.lastname: Okuda
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Mizukami
authors.forename: Keiya
authors.initials: K
authors.lastname: Ozawa
authors.forename: Tomoyuki
authors.initials: T
authors.lastname: Saito
mesh_terms: 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
chemical_list: D005819:Genetic Markers; D001616:beta-Galactosidase