Ex vivo expansion and pluripotential differentiation of cryopreserved human bone marrow mesenchymal stem cells |
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Authors: | Xiang Ying Zheng Qiang Jia Bing-bing Huang Guo-ping Xu Yu-lin Wang Jin-fu Pan Zhi-jun |
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Institution: | (1) School of Life Sciences, Zhejiang University, Hangzhou, 310012, China;(2) Department of Orthopedics, the Second Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou, 310006, China |
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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.
Project supported by the Science Foundation (No. 2003C23015) and the Natural Science Foundation (No Y204139) of Zhejiang Province,
China |
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Keywords: | Bone marrow mesenchymal stem cells Cryopreservation Expansion Differentiation |
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