Chinese Journal of Tissue Engineering Research ›› 2010, Vol. 14 ›› Issue (23): 4217-4221.doi: 10.3969/j.issn.1673-8225.2010.23.009

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Transforming growth factor-beta 1/ insulin-like growth factor-1 versus nucleus pulposus cells in inducing the differentiation of adipose-derived mesenchymal stem cells into nucleus pulposus-like cells

Liu Lian-jiang1,2, Li Fang2, Wen Tian-yong1   

  1. 1Fourth Military Medical University of Chinese PLA, Xi’an  710032, Shaanxi Province, China;
    2Orthopaedic Research Institute, General Hospital of Beijing Military Area Command of Chinese PLA, Beijing  100700, China
  • Online:2010-06-04 Published:2010-06-04
  • Contact: Li Fang, Professor, Master’s supervisor, Orthopaedic Research Institute, General Hospital of Beijing Military Area Command of Chinese PLA, Beijing 100700, China FANGL6722@sina. com
  • About author:Liu Lian-jiang, Studying for master’s degree, Physician, Fourth Military Medical University of Chinese PLA, Xi’an 710032, Shaanxi Province, China; Orthopaedic Research Institute, General Hospital of Beijing Military Area Command of Chinese PLA, Beijing 100700, China woodl@126.com
  • Supported by:

    the Natural Science Foundation of Beijing City, No. 5062039*

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Abstract:

BACKGROUND: Previous studies have addressed adipose-derived mesenchymal stem cells (ADSCs) culture way for monolayer culture; induction way was mainly cytokine method, and under intervertebral discs micro-environment, cell interactions also deserved further study in its three-dimensional environment, in addition to the effects on cell cytokines.

OBJECTIVE: To observe differences in differentiation of ADSCs into NPCs under two induced conditions: nucleus pulposus cells (NPCs) and transforming growth factor β1 (TGF-β1)/ insulin-like growth factor (IGF-1) in vitro.

METHODS: Rabbit ADSCs monolayer culture and NPCs were cultured respectively, 3 generations of ADSCs were qualified; 5×106 cells were made into microspheres and placed in Transwell culture plates for culture under TGF-β 1/IGF-1 induction. Cell shape changes were observed before, 7, 14 days after induction. Reverse transcription-polymerase chain reaction (RT-PCR) was used to determine levels of type II collagen and proteoglycan.

RESULTS AND CONCLUSION: At 7 and 14 days, no significant difference was detected in size and shape of ADSCs microspheres in both groups. RT-PCR detection results displayed that type II collagen and proteoglycan mRNA expression was determined in 7-day TGF-β1/IGF-1 group and NPCs-induced group, but expression was stronger in the TGF-β1/IGF-1 group. Type II collagen and proteoglycan mRNA expression was obviously increased in 14-day NPCs-induced group, superior to TGF-β1/IGF-1 group. Results have suggested that in vitro NPCs and TGF-β1/IGF-1 lead to the ADSCs, which had promoting effects on NPCs differentiation. TGF-β 1 and IGF-1 are normal NPCs-secreted important factors. The promoting effects on proliferation and differentiation exist, with the exception of cytokine induction effect, in differentiation of ADSCs into NPCs.

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