title: Nonviral gene transfer into human meniscal cells. Part II: effect of three-dimensional environment and overexpression of human fibroblast growth factor 2.
abstract: OBJECTIVE
  Our aim was to study the effect of three-dimensional (3D) environment and overexpression of human fibroblast growth factor 2 (FGF-2) on meniscal fibrochondrocytes in vitro.
  
  
  METHODS
  Human meniscal fibrochondrocytes were transfected with expression plasmid vectors carrying the Photinus pyralis luciferase gene, the Escherichia coli β-galactosidase gene or a human FGF-2 cDNA. Modified fibrochondrocytes were cultivated in 3D alginate hydrogel or cell pellets or in 2D monolayer culture.
  
  
  RESULTS
  The levels of luciferase activity showed a peak at day two and returned to baseline levels by day 11, regardless of the type of cultivation. Both 3D environments supported the secretion of human FGF-2 protein upon FGF-2 transfection. Overexpression of human FGF-2 by genetically modified human meniscal fibrochondrocytes stimulated proliferation but not glycosaminoglycan synthesis only in 3D culture. Culture in alginate spheres resulted in a larger difference in cell numbers compared with pellet cultures.
  
  
  CONCLUSIONS
  Three-dimensional alginate spheres are well suited for the culture of genetically modified human meniscal fibrochondrocytes. These data are of value for cell-based approaches to meniscal repair using genetically modified human meniscal fibrochondrocytes overexpressing human FGF-2.
authors: Center of Experimental Orthopaedics, Saarland University, 66421, Homburg, Germany.
authors: Hsiao-Ping
authors: HP
authors: Lee
authors: Ana
authors: A
authors: Rey-Rico
authors: Magali
authors: M
authors: Cucchiarini
authors: Henning
authors: H
authors: Madry
mesh_terms: D000464:Alginates; D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D002470:Cell Survival; D017690:Cell Transplantation / Q000379:methods*; D002478:Cells, Cultured; D019902:Chondrocytes / Q000166:cytology / Q000378:metabolism / Q000637:transplantation*; D004926:Escherichia coli / Q000235:genetics; D016222:Fibroblast Growth Factor 2 / Q000235:genetics / Q000378:metabolism*; D048888:Fireflies / Q000235:genetics; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors; D020723:Glucuronic Acid; D006603:Hexuronic Acids; D006801:Humans; D066298:In Vitro Techniques; D008592:Menisci, Tibial / Q000166:cytology / Q000378:metabolism / Q000601:surgery*; D015854:Up-Regulation
chemical_list: D000464:Alginates; D006603:Hexuronic Acids; D016222:Fibroblast Growth Factor 2; D020723:Glucuronic Acid