title: [CHONDROGENESIS-SPECIFIC MICRORNA EXPRESSION PATTERN ANALYSIS IN CHONDROGENIC DIFFERENTIATION OF HUMAN ADIPOSE-DERIVED STEM CELLS].
abstract: OBJECTIVE
  To investigate the microRNA (miRNA) expression profile during chondrogenic differentiation of human adipose-derived stem cells (hADSCs), and assess the roles of involved miRNAs during chondrogenesis.
  
  
  METHODS
  hADSCs were harvested and cultured from donors who underwent elective liposuction or other abdominal surgery. When the cells were passaged to P3, chondrogenic induction medium was used for chondrogenic differentiation. The morphology of the cells was observed by inverted phase contrast microscopy. Alcian blue staining was carried out at 21 days after induction to access the chondrogenic status. The expressions of chondrogenic proteins were detected by ELISA at 0, 7, 14, and 21 days. The miRNA expression profiles at pre- and post-chondrogenic induction were obtained by microarray assay, and differentially expressed miRNAs were verified by real-time quantitative PCR (qRT-PCR). The targets of the miRNAs were predicted by online software programs.
  
  
  RESULTS
  hADSCs were cultured successfully and induced with chondrogenic medium. At 21 days after chondrogenic induction, the cells were stained positively for alcian blue staining. At 7, 14, and 21 days after chondrogenic induction, the levels of collogen type II, Col2a1, aggrecan, Coll0a1, and chondroitin sulfate in induced hADSCs were significantly higher than those in non-induced hADSCs (P<0.05). Eleven differentially expressed miRNAs were found, including seven up-regulated and four down-regulated. Predicted target genes of the differentially expressed miRNAs were based on the overlap from three public prediction algorithms, with the known functions of regulating chondrogenic differentiation of stem cells, self-renewal, signal transduction, intracellular signaling cascade, and cell cycle control.
  
  
  CONCLUSIONS
  A group of miRNAs and their target genes are identified, which may play important roles in regulating chondrogenic differentiation of hADSCs. These results will facilitate the initial understanding of the molecular mechanism of chondrogenic differentiation in hADSCs and subsequently control hADSCs differentiation, and provide high performance seed cells for cartilage tissue engineering.
authors.forename: Ziji
authors.initials: Z
authors.lastname: Zhang
authors.forename: Yan
authors.initials: Y
authors.lastname: Kang
authors.forename: Zhiqi
authors.initials: Z
authors.lastname: Zhang
authors.forename: Zibo
authors.initials: Z
authors.lastname: Yang
authors.forename: Shuying
authors.initials: S
authors.lastname: Fang
authors.forename: Puyi
authors.initials: P
authors.lastname: Sheng
authors.forename: Aishan
authors.initials: A
authors.lastname: He
authors.forename: Ming
authors.initials: M
authors.lastname: Fu
authors.forename: Weiming
authors.initials: W
authors.lastname: Liao
mesh_terms: D017667:Adipocytes / Q000378:metabolism*; D000368:Aged; D053674:Aggrecans; D002356:Cartilage; D002454:Cell Differentiation / Q000235:genetics*; D002478:Cells, Cultured; D020219:Chondrogenesis / Q000235:genetics*; D002809:Chondroitin Sulfates; D024043:Collagen Type II; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D035683:MicroRNAs / Q000235:genetics*; D013234:Stem Cells; D023822:Tissue Engineering
chemical_list: D053674:Aggrecans; D024043:Collagen Type II; D035683:MicroRNAs; D002809:Chondroitin Sulfates