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Differentiation of mesenchymal stem cells towards a nucleus pulposus-like phenotype utilizing simulated microgravity In vitro

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Summary

Mesenchymal stem cells (MSCs) were induced into a nucleus pulposus-like phenotype utilizing simulated microgravity in vitro in order to establish a new cell-based tissue engineering treatment for intervertebral disc degeneration. For induction of a nucleus pulposus-like phenotype, MSCs were cultured in simulated microgravity in a chemically defined medium supplemented with 0 (experimental group) and 10 ng/mL (positive control group) of transforming growth factor β1 (TGF-β1). MSCs cultured under conventional condition without TGF-β1 served as blank control group. On the day 3 of culture, cellular proliferation was determined by WST-8 assay. Differentiation markers were evaluated by histology and reverse transcriptase-polymerase chain reaction (RT-PCR). TGF-β1 slightly promoted the proliferation of MSCs. The collagen and proteoglycans were detected in both groups after culture for 7 days. The accumulation of proteoglycans was markedly increased. The RT-PCR revealed that the gene expression of Sox-9, aggrecan and type II collagen, which were chondrocyte specific, was increased in MSCs cultured under simulated microgravity for 3 days. The ratio of proteoglycans/collagen in blank control group was 3.4-fold higher than positive control group, which denoted a nucleus pulposus-like phenotype differentiation. Independent, spontaneous differentiation of MSCs towards a nucleus pulposus-like phenotype in simulated microgravity occurred without addition of any external bioactive stimulators, namely factors from TGF-β family, which were previously considered necessary.

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Authors and Affiliations

  1. Department of Orthopedics, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, China

    Wei Luo  (罗 伟), Wei Xiong  (熊 伟), Min Qiu  (邱 敏), Yongwei Lv  (吕永威), Yong Li  (李 勇) & Feng Li  (李 锋)

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  1. Wei Luo  (罗 伟)
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  2. Wei Xiong  (熊 伟)
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  3. Min Qiu  (邱 敏)
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  4. Yongwei Lv  (吕永威)
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  5. Yong Li  (李 勇)
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  6. Feng Li  (李 锋)
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Corresponding author

Correspondence to Wei Xiong  (熊 伟).

Additional information

This project was supported by grants from the National Natural Sciences Foundation of China (No. 30772206 & 10925208).

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Luo, W., Xiong, W., Qiu, M. et al. Differentiation of mesenchymal stem cells towards a nucleus pulposus-like phenotype utilizing simulated microgravity In vitro . J. Huazhong Univ. Sci. Technol. [Med. Sci.] 31, 199–203 (2011). https://doi.org/10.1007/s11596-011-0252-3

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  • DOI: https://doi.org/10.1007/s11596-011-0252-3

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