PMID- 23253807
OWN - NLM
STAT- MEDLINE
DCOM- 20130905
LR  - 20181202
IS  - 0376-2491 (Print)
IS  - 0376-2491 (Linking)
VI  - 92
IP  - 29
DP  - 2012 Aug 7
TI  - [SOX9 enhanced chondrogenic differentiation potential of human umbilical cord 
      mesenchymal stem cells through cellular aggregation].
PG  - 2050-4
AB  - OBJECTIVE: To explore the effects of transcription factor SOX9 on chondrogenic 
      differentiation potential of human umbilical cord mesenchymal stem cells 
      (hUC-MSCs). METHODS: hUC-MSCs were harvested from human umbilical cord and their 
      phenotypic characteristics identified by flow cytometry. To confirm their 
      multipotency, hUC-MSCs were induced to differentiate toward adiposity and 
      osteogenesis. After transfection with the packaging lentivirus vectors containing 
      SOX9 in vitro, the expression of green fluorescent protein (GFP) and the 
      efficiency of transfection were detected by fluorescence microscopy. Their 
      cellular proliferation capacity was detected by thiazolyl blue tetrazolium 
      bromide (MTT) assay.hUC-MSCs modified with SOX9 were seeded into monolayer and 
      cultured for 21 days in a defined, serum-free medium supplemented with 
      transforming growth factor (TGF)-beta1. The untransduced cells or those transduced 
      with GFP served as the controls. Morphologic changes of hUC-MSCs were observed 
      daily and their chondrogenic differentiation was evaluated by reverse 
      transcription-polymerase chain reaction (RT-PCR), Western blot, and 
      immunofluorescent staining. And the accumulation of sulfated glycosaminoglycans 
      was detected by Alcian blue staining. Meanwhile, the expressions of collagen I, X 
      and cell adhesion molecule N-cadherin were assayed. RESULTS: The hUC-MSCs 
      isolated from human umbilical cord stromas exhibited fibroblastic morphology and 
      they were positive for CD29 (95.9%), CD44 (96.5%), CD90 (98.9%), CD105 (94.3%) 
      and negative for hematopoietic stem cells surface markers CD34 (3.0%) and CD45 
      (2.6%). At Day 21, hUC-MSCs differentiated toward adiposity and osteogenesis. 
      Both oil red O and alkaline phosphatase stains were intensely positive and it 
      confirmed the multilineage potential of hUC-MSCs. An intense expression of GFP 
      was observed under flourescence microscope and the transfection efficiency of 
      cells with Lenti-GFP-SOX9 or Lenti-GFP was more than 90% respectively. SOX9 gene 
      was over-expressed in hUC-MSCs at 48 h post-transduction. The proliferation of 
      hUC-MSCs had no significant effect after the transfection of lentivirus vectors 
      (P > 0.05). In vitro high-density monolayer culture of these SOX9-transfected 
      hUC-MSCs demonstrated that spontaneous cell aggregation appeared at Day 14 of 
      culturing and subsequently generated large cartilage nodules. However there was 
      no phenomenon of cell aggregation occurring in the cells transducted by Lenti-GFP 
      or untransduced vectors. The expressions of collagen II and Aggrecan were higher 
      in SOX9 transducted cells than those in the controls. Alcian blue staining also 
      showed abundant accumulation of sulfated glycosaminoglycans in the SOX9-induced 
      cartilage nodules. The expression of collagen I had no difference in all groups 
      and collagen X was inhibited in SOX9 transduced cells. N-cadherin was strongly 
      up-regulated by SOX9 and might result in cellular aggregation and formation of 
      large cartilage nodules. CONCLUSION: SOX9 may enhance the chondrogenic 
      differentiation potential of human umbilical cord mesenchymal stem cells through 
      cellular aggregation.
FAU - Xu, Yun
AU  - Xu Y
AD  - Department of Orthopedics, Xiangcheng People's Hospital of Suzhou, Suzhou, China.
FAU - Chen, Liang
AU  - Chen L
FAU - Shi, Yong
AU  - Shi Y
FAU - Gu, Yong
AU  - Gu Y
FAU - Zou, Jun
AU  - Zou J
FAU - Huang, Chen
AU  - Huang C
FAU - Tang, Tian-si
AU  - Tang TS
LA  - chi
PT  - Journal Article
PL  - China
TA  - Zhonghua Yi Xue Za Zhi
JT  - Zhonghua yi xue za zhi
JID - 7511141
RN  - 0 (SOX9 Transcription Factor)
RN  - 0 (SOX9 protein, human)
SB  - IM
MH  - Cell Aggregation
MH  - Cell Differentiation
MH  - Cells, Cultured
MH  - Chondrocytes/*cytology
MH  - *Chondrogenesis
MH  - Flow Cytometry
MH  - Humans
MH  - Mesenchymal Stem Cells/*cytology
MH  - SOX9 Transcription Factor/*genetics
MH  - Transfection
MH  - Umbilical Cord/cytology
EDAT- 2012/12/21 06:00
MHDA- 2013/09/06 06:00
CRDT- 2012/12/21 06:00
PHST- 2012/12/21 06:00 [entrez]
PHST- 2012/12/21 06:00 [pubmed]
PHST- 2013/09/06 06:00 [medline]
PST - ppublish
SO  - Zhonghua Yi Xue Za Zhi. 2012 Aug 7;92(29):2050-4.