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Ex vivo expansion and pluripotential differentiation of cryopreserved human bone marrow mesenchymal stem cells

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Abstract

This study is aimed at investigating the potentials of ex vivo expansion and pluri-differentiation of cryopreservation of adult human bone marrow mesenchymal stem cells (hMSCs) into chondrocytes, adipocytes and neurocytes. Cryopreserved hMSCs were resuscitated and cultured for 15 passages, and then induced into chondrocytes, adipocytes and neurocytes with corresponding induction medium. The induced cells were observed for morphological properties and detected for expressions of type II collagen, triglyceride or neuron-specific enolase and nestin. The result showed that the resuscitated cells could differentiate into chondrocytes after exposure to transforming growth factor β1 (TGF-β1), insulin-like growth factor I (IGF-I) and vitamin C (VC), and uniformly changed morphologically from a spindle-like fibroblastic appearance to a polygonal shape in three weeks. The induced cells were heterochromatic to safranin O and expressed cartilage matrix-procollagenal (II) mRNA. The resuscitated cells cultured in induction medium consisting of dexamethasone, 3-isobutyl-1-methylxanthine, indomethacin and IGF-I showed adipogenesis, and lipid vacuoles accumulation was detectable after 21 d. The resuscitated hMSCs were also induced into neurocytes and expressed nestin and neuron specific endolase (NSE) that were special surface markers associated with neural cells at different stage. This study suggested that the resuscitated hMSCs should be still a population of pluripotential cells and that it could be used for establishing an abundant hMSC reservoir for further experiment and treatment of various clinical diseases.

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Authors and Affiliations

  1. School of Life Sciences, Zhejiang University, Hangzhou, 310012, China

    Xiang Ying, Jia Bing-bing, Huang Guo-ping, Xu Yu-lin & Wang Jin-fu

  2. Department of Orthopedics, the Second Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou, 310006, China

    Zheng Qiang & Pan Zhi-jun

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  1. Xiang Ying
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  2. Zheng Qiang
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  3. Jia Bing-bing
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  4. Huang Guo-ping
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  5. Xu Yu-lin
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  6. Wang Jin-fu
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  7. Pan Zhi-jun
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Correspondence to Wang Jin-fu or Pan Zhi-jun.

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Project supported by the Science Foundation (No. 2003C23015) and the Natural Science Foundation (No Y204139) of Zhejiang Province, China

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Xiang, Y., Zheng, Q., Jia, Bb. et al. Ex vivo expansion and pluripotential differentiation of cryopreserved human bone marrow mesenchymal stem cells. J. Zhejiang Univ. - Sci. B 8, 136–146 (2007). https://doi.org/10.1631/jzus.2007.B0136

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  • DOI: https://doi.org/10.1631/jzus.2007.B0136

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