title: Characterization of Chondrogenic Gene Expression and Cartilage Phenotype Differentiation in Human Breast Adipose-Derived Stem Cells Promoted by Ginsenoside Rg1 In Vitro.
abstract: OBJECTIVE
  Investigating and understanding chondrogenic gene expression during the differentiation of human breast adipose-derived stem cells (HBASCs) into chondrogenic cells is a prerequisite for the application of this approach for cartilage repair and regeneration. In this study, we aim to characterize HBASCs and to examine chondrogenic gene expression in chondrogenic inductive culture medium containing ginsenoside Rg1.
  
  
  METHODS
  Human breast adipose-derived stem cells at passage 3 were evaluated based on specific cell markers and their multilineage differentiation capacity. Cultured HBASCs were treated either with basic chondrogenic inductive conditioned medium alone (group A, control) or with basic chondrogenic inductive medium plus 10 µg/ml (group B), 50 µg/ml (group C), or 100µg/ml ginsenoside Rg1 (group D). Cell proliferation was assessed using the CCK-8 assay for a period of 9 days. Two weeks after induction, the expression of chondrogenic genes (collagen type II, collagen type XI, ACP, COMP and ELASTIN) was determined using real-time PCR in all groups.
  
  
  RESULTS
  The different concentrations of ginsenoside Rg1 that were added to the basic chondrogenic inductive culture medium promoted the proliferation of HBASCs at earlier stages (groups B, C, and D) but resulted in chondrogenic phenotype differentiation and higher mRNA expression of collagen type II (CO-II), collagen type XI (CO-XI), acid phosphatase (ACP), cartilage oligomeric matrix protein (COMP) and ELASTIN compared with the control (group A) at later stages. The results reveal an obvious positive dose-effect relationship between ginsenoside Rg1 and the proliferation and chondrogenic phenotype differentiation of HBASCs in vitro.
  
  
  CONCLUSIONS
  Human breast adipose-derived stem cells retain stem cell characteristics after expansion in culture through passage 3 and serve as a feasible source of cells for cartilage regeneration in vitro. Chondrogenesis in HBASCs was found to be prominent after chondrogenic induction in conditions containing ginsenoside Rg1.
authors.forename: Fang-Tian
authors.initials: FT
authors.lastname: Xu
authors.forename: Hong-Mian
authors.initials: HM
authors.lastname: Li
authors.forename: Cheng-Yi
authors.initials: CY
authors.lastname: Zhao
authors.forename: Zhi-Jie
authors.initials: ZJ
authors.lastname: Liang
authors.forename: Min-Hong
authors.initials: MH
authors.lastname: Huang
authors.forename: Qing
authors.initials: Q
authors.lastname: Li
authors.forename: Ying-Chao
authors.initials: YC
authors.lastname: Chen
authors.forename: Gang-Yi
authors.initials: GY
authors.lastname: Chi
mesh_terms: D000135:Acid Phosphatase / Q000235:genetics / Q000378:metabolism; D000273:Adipose Tissue / Q000166:cytology; D015703:Antigens, CD / Q000378:metabolism; D001940:Breast / Q000166:cytology; D002356:Cartilage / Q000378:metabolism; D064236:Cartilage Oligomeric Matrix Protein / Q000235:genetics / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects*; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D019902:Chondrocytes / Q000166:cytology / Q000378:metabolism*; D020219:Chondrogenesis / Q000187:drug effects*; D024043:Collagen Type II / Q000235:genetics / Q000378:metabolism; D024081:Collagen Type IX / Q000235:genetics / Q000378:metabolism; D017077:Culture Media, Conditioned / Q000494:pharmacology; D004549:Elastin / Q000235:genetics / Q000378:metabolism; D005260:Female; D036145:Ginsenosides / Q000494:pharmacology*; D006801:Humans; D016130:Immunophenotyping; D060888:Real-Time Polymerase Chain Reaction; D013234:Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism
chemical_list: D015703:Antigens, CD; D064236:Cartilage Oligomeric Matrix Protein; D024043:Collagen Type II; D024081:Collagen Type IX; D017077:Culture Media, Conditioned; D036145:Ginsenosides; D004549:Elastin; D000135:Acid Phosphatase; C035054:ginsenoside Rg1