title: [Experimental study on adipose-derived stem cells transfected by bone morphogenetic protein 14 co-culture with chondrocytes].
abstract: OBJECTIVE
  To evaluate the synergistic effect of bone morphogenetic protein 14 (BMP-14) and chondrocytes co-culture on chondrogenesis of adipose-derived stem cells (ADSCs) so as to optimize the source of seed cells for cartilage tissue engineering.
  
  
  METHODS
  ADSCs and chondrocytes were isolated and cultured respectively from articular cartilage and subcutaneous fat of 2 male New Zealand white rabbits (weighing, 1.5 kg and 2.0 kg). The cells at passage 3 were harvested for experiment. ADSCs were identified by osteogenic induction (alizarin red staining), chondrogenic induction (alcian blue staining), and adipogenic induction (oil red O staining). The optimum multiplicity of infection (MOI) of transfection of adenovirus-cytomegalovirus (CMV)-BMP-14-internal ribosome entry site (IRES)-human renilla reniformis green fluorescent protein 1 (hrGFP-1) was determined and then ADSCs were transfected by the optimum MOI. The experiment was divided into 5 groups: group A, co-culture of ADSCs transfected by BMP-14 and chondrocytes (1:1 in Transwell chambers); group B, co-culture of ADSCs and chondrocytes (1:1 in Transwell chambers); group C, culture of ADSCs transfected by BMP-14; group D, simple chondrocytes culture; and group E, simple ADSCs culture. After 3 weeks, the glycosaminoglycan (GAG) content was detected by alcian blue staining; the expressions of collagen type II and BMP-14 protein were detected by Western blot; expression of Sox-9 gene was detected by RT-PCR.
  
  
  RESULTS
  The cultured cells were proved to be ADSCs by identification. Inverted fluorescence microscope showed optimum transfection effect when MOI was 150. GAG content, expressions of collagen type II and BMP-14 protein, expression of Sox-9 gene were significantly higher in groups A and C than in the other 3 groups, in group A than in group C (P < 0.05), and groups B and D were significantly higher than group E (P < 0.05), but no significant difference was found between groups B and D (P > 0.05).
  
  
  CONCLUSIONS
  It can promote differentiation of ADSCs into chondrocytes by BMP-14 co-culture with chondrocytes, and they have a synergistic effect.
authors.affiliation: Department of Burn and Plastic Surgery, the First Affiliated Hospital of Liaoning Medical College, Jinzhou Liaoning, 121001, P.R. China.
authors.forename: Hong
authors.initials: H
authors.lastname: Yuan
authors.forename: Jieyuan
authors.initials: J
authors.lastname: Zhang
authors.forename: Rongming
authors.initials: R
authors.lastname: Zhang
mesh_terms: D000273:Adipose Tissue / Q000166:cytology*; D000818:Animals; D019485:Bone Morphogenetic Proteins / Q000235:genetics* / Q000378:metabolism; D002358:Cartilage, Articular / Q000166:cytology; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation; D002478:Cells, Cultured; D019902:Chondrocytes / Q000166:cytology* / Q000378:metabolism; D018920:Coculture Techniques; D024043:Collagen Type II / Q000378:metabolism; D003587:Cytomegalovirus / Q000235:genetics; D006025:Glycosaminoglycans / Q000378:metabolism; D008297:Male; D011817:Rabbits; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism; D023822:Tissue Engineering / Q000379:methods*; D014162:Transfection
chemical_list: D019485:Bone Morphogenetic Proteins; D024043:Collagen Type II; D006025:Glycosaminoglycans