title: Nonviral gene transfer to human meniscal cells. Part I: transfection analyses and cell transplantation to meniscus explants.
abstract: OBJECTIVE
  Our aim was to evaluate whether nonviral vectors can genetically modify primary human juvenile and adult meniscal fibrochondrocytes at low toxicity in vitro and to test the hypothesis that transfected human meniscal fibrochondrocytes transplanted into longitudinal defects and onto human medial meniscus explant cultures are capable of expressing transgene products in vitro.
  
  
  METHODS
  Eighteen nonviral gene transfer systems were examined to identify the best suited method for an efficient transfection of primary cultures of juvenile and adult human meniscal fibrochondrocytes using luciferase and lacZ reporter gene constructs and then transplanted to meniscus explant cultures.
  
  
  RESULTS
  Gene transfer systems FuGENE 6, GeneJammer, TurboFectin 8, calcium phosphate co-precipitates and GeneJuice led to minimal toxicity in both cell types. Nanofectin 2 and JetPEI resulted in maximal luciferase activity in both cell types. Maximal transfection efficiency based on X-gal staining following lacZ gene transfer was achieved using Lipofectamine 2000, revealing a mean transfection efficiency of 8.6 % in human juvenile and of 8.4 % in adult meniscal fibrochondrocytes. Transfected, transplanted meniscal fibrochondrocytes adhered to the meniscal tissue and continued to express the transgene for at least five days following transfection.
  
  
  CONCLUSIONS
  Nonviral gene transfer systems are safe and capable of transfecting both juvenile and adult human meniscal fibrochondrocytes, which, when transplanted to meniscal tissue in vitro, permit the expression of selected transgenes to be maintained. These results are of value for combining gene therapy and cell transplantation approaches as a means to enhance meniscal repair.
authors: Center of Experimental Orthopaedics, Saarland University, 66421, Homburg, Germany.
authors: Hsiao-Ping
authors: HP
authors: Lee
authors: Gunter
authors: G
authors: Kaul
authors: Magali
authors: M
authors: Cucchiarini
authors: Henning
authors: H
authors: Madry
mesh_terms: D000367:Age Factors; D002470:Cell Survival; D017690:Cell Transplantation / Q000379:methods*; D002478:Cells, Cultured; D019902:Chondrocytes / Q000166:cytology / Q000637:transplantation*; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors; D006801:Humans; D066298:In Vitro Techniques; D008592:Menisci, Tibial / Q000166:cytology / Q000601:surgery*; D014162:Transfection / Q000379:methods*