title: Ex vivo expansion and pluripotential differentiation of cryopreserved human bone marrow mesenchymal stem cells.
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 beta(1) (TGF-beta(1)), insulin-like growth factor I (IGF-I) and vitamin C (V(C)), 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.
authors.affiliation: School of Life Sciences, Zhejiang University, Hangzhou, China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Xiang
authors.forename: Qiang
authors.initials: Q
authors.lastname: Zheng
authors.forename: Bing-bing
authors.initials: BB
authors.lastname: Jia
authors.forename: Guo-ping
authors.initials: GP
authors.lastname: Huang
authors.forename: Yu-lin
authors.initials: YL
authors.lastname: Xu
authors.forename: Jin-fu
authors.initials: JF
authors.lastname: Wang
authors.forename: Zhi-jun
authors.initials: ZJ
authors.lastname: Pan
mesh_terms: D001854:Bone Marrow Cells / Q000166:cytology*; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation; D002478:Cells, Cultured; D015925:Cryopreservation / Q000379:methods*; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D039904:Pluripotent Stem Cells / Q000166:cytology*; D023822:Tissue Engineering / Q000379:methods*