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Characterization and gene transfer in mesenchymal stem cells derived from human umbilical-cord blood
Authors:Lu Fei-Zhou  Fujino Masayuki  Kitazawa Yusuke  Uyama Taro  Hara Yuko  Funeshima Naoko  Jiang Jian-Yuan  Umezawa Akihiro  Li Xiao-Kang
Affiliation:Laboratory of Transplantation Immunology, National Research Institute for Child Health and Development, Tokyo, Japan.
Abstract:It has been shown that the stromal-cell population found in bone marrow can be expanded and differentiated into cells with the phenotypes of bone, cartilage, muscle, neural, and fat cells. However, whether mesenchymal stem cells (MSCs) are present in human umbilical-cord blood (UCB) has been the subject of ongoing debate. In this study, we report on a population of fibroblastlike cells derived from the mononuclear fraction of human UCB with osteogenic and adipogenic potential, as well as the presence of a subset of cells that have been maintained in continuous culture for more than 6 months. These cells were found to express CD29, CD44, CD90, CD95, CD105, CD166, and MHC class, but not CD14, CD34, CD40, CD45, CD80, CD86, CD117, CD152, or MHC class II. We also compared gene expression after gene transfer using lenti- and adenoviral vectors carrying the green fluorescence protein to the MSCs derived from UCB because a reliable gene-delivery system is required to transfer target genes into MSCs, which have attracted attention as potential platforms for the systemic delivery of therapeutic genes. The lentiviral vectors can transduce these cells more efficiently than can adenoviral vectors, and we maintained transgene expression for at least 5 weeks. This is the first report showing that UCB-derived MSCs can express exogenous genes by way of a lentivirus vector. These results demonstrate that human UCB is a source of mesenchymal progenitors and may be used in cell transplantation and a wide range of gene-therapy treatments.
Keywords:adeno-GFP, recombinant adenovirus expressing the green fluorescent protein gene Ax1CAGFP   BBS, BBS 50 mmol/L BES, 280 mmol/L NaCl, 1.5 mmol/L Na2HPO4   BM, bone marrow   DMEM, Dulbecco’s modified Eagle medium   EDTA, ethylenediaminetetraacetic acid   FACS, fluorescence-activated cell sorting   FBS, fetal bovine serum   FITC, fluorescein isothiocyanate   GFP, green fluorescent protein   IU, infectious unit   lenti-GFP, lentivirus expressing the green fluorescent protein   MNC, mononuclear cell   MSC, mesenchymal stem cell   MSCGM, MSC growth medium   PBS, phosphate-buffered saline solution   TBST, Tris-buffered saline solution containing Tween-20   UCB, umbilical-cord blood
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