title: Differentiation of immortalized human precartilaginous stem cells into nucleus pulposus-like cells.
abstract: OBJECTIVE
  We aimed to explore the differentiation of immortalized human precartilaginous stem cells (IPSCs) into nucleus pulposus (NP)-like cells induced by transforming growth factor-β1 (TGF-β1) and examine its biological characteristics.
  
  
  METHODS
  The IPSCs were seeded onto chitosan/glycerophosphate (C/GP) scaffolds and induced into NP-like cells by adding TGF-β1 under hypoxic conditions. The growth and differentiation of IPSCs were observed, and the formation of glycosaminoglycans (GAG) in the extracellular matrix of differentiating cells was detected by Alcian Blue staining. The expressions of type II collagen and aggrecan genes in NP-like cells were examined by reverse transcription-polymerase chain reaction (RT-PCR). The expressions of Sox9 and β-catenin were analyzed by Western blotting.
  
  
  RESULTS
  The IPSCs were observed to grow well on the C/GP scaffolds. After 7 days, Alcian Blue staining demonstrated the formation of GAG. The RT-PCR results showed that expression of type II collagen and aggrecan were upregulated compared with control group (P<0.05, P<0.05). Likewise, western blotting results showed that the expression of Sox9 and β-catenin was upregulated compared with control group (P<0.05, P<0.05).
  
  
  CONCLUSIONS
  The findings demonstrate that IPSCs could be differentiated into NP-like cells following induction by TGF-β1.
authors.affiliation: Department of Orthopedics, Wuxi People's Hospital, Nanjing Medical University Wuxi 214023, China.
authors.forename: Qiong
authors.initials: Q
authors.lastname: Wang
authors.affiliation: Department of Orthopedics, Wuxi People's Hospital, Nanjing Medical University Wuxi 214023, China.
authors.forename: Xiaofeng
authors.initials: X
authors.lastname: Gu
authors.affiliation: Department of Orthopedics, Wuxi People's Hospital, Nanjing Medical University Wuxi 214023, China.
authors.forename: Li
authors.initials: L
authors.lastname: Cheng
authors.affiliation: Department of Orthopedics, Wuxi People's Hospital, Nanjing Medical University Wuxi 214023, China.
authors.forename: Junfang
authors.initials: J
authors.lastname: Wang
mesh_terms: D053674:Aggrecans / Q000235:genetics / Q000378:metabolism; D015153:Blotting, Western; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000187:drug effects; D015687:Cell Hypoxia; D002460:Cell Line; D019902:Chondrocytes / Q000187:drug effects / Q000378:metabolism / Q000502:physiology*; D024043:Collagen Type II / Q000235:genetics / Q000378:metabolism; D018507:Gene Expression Regulation, Developmental; D006025:Glycosaminoglycans / Q000378:metabolism; D006801:Humans; D007403:Intervertebral Disc / Q000166:cytology*; D010641:Phenotype; D020133:Reverse Transcriptase Polymerase Chain Reaction; D055758:SOX9 Transcription Factor / Q000378:metabolism; D013234:Stem Cells / Q000187:drug effects / Q000378:metabolism / Q000502:physiology*; D013997:Time Factors; D054457:Tissue Scaffolds; D053773:Transforming Growth Factor beta1 / Q000494:pharmacology; D051176:beta Catenin / Q000378:metabolism
chemical_list: D053674:Aggrecans; C495270:CTNNB1 protein, human; D024043:Collagen Type II; D006025:Glycosaminoglycans; D055758:SOX9 Transcription Factor; C527932:SOX9 protein, human; D053773:Transforming Growth Factor beta1; D051176:beta Catenin
keywords: Precartilaginous stem cells (IPSCs); nucleus pulposus-liked cells; transforming growth factor-β1