title: [The effect of transgenic cell strains expressing recombinant adenovirus vector of hOSM gene on the proliferation and homing ability of umbilical cord blood CD34(+) hematopoietic stem/progenitor cells].
abstract: OBJECTIVE
  To establish the transgenic cell strains expressing recombinant adenovirus vector of human Oncostain M(hOSM)gene which is supposed to be used as feeder layer cells for the proliferation of umbilical cord blood CD34(+) hematopoietic stem/progenitor cell (HSPC) and compare its migration capacity before and after amplification in vitro.
  
  
  METHODS
  Establish the transgenic cell strains expressing recombinant adenovirus vector of hOSM gene, and the objective gene was detected by RT-PCR and ELISA. The purity of umbilical cord blood CD34(+) HSPC separated by magnetic-activated cell sorting (MACS) was detected by the FCM. After culturing with feeder layer cells, detect the rate of proliferation by flow cytometry (FCM). To compare the homing ability of HSPC after amplification in vitro, detect the spontaneous migration rate and migration rate induced by SDF-1 using transmembrane migration assay (Transwell experiment).
  
  
  RESULTS
  The green fluorescence was observed by fluorescence microscope in the transgenic cell strains, and the objective gene was confirmed by RT-PCR and ELISA.The purity of umbilical cord blood CD34(+) HSPC separated by MACS could reach(96.8 ± 2.28)%. After culturing with feeder layer cells for 7 days, the CD34(+) cells were 15.73 times in group containing hOSM more than in group without hOSM. The expression rate of adhension molecules on the surface of CD34(+) cells were also higher in the group containing hOSM than without hOSM. After using Transwell assys to detect the homing ability of culturing cells, the induction migration rate of stem cells clturing on transgenic cell strains was (40.68 ± 1.35)%, significantly higher than the control, which reveals a better homing ability.
  
  
  CONCLUSIONS
  Recombinant adenovirus vector of hOSM gene as feeder layer cells can effectively proliferate umbilical cord blood CD34(+) HSPC in vitro and delay it differentiate, what's more, the stem cells retain a high homing ability after culturing on transgenic cell strains in vitro.
authors.affiliation: Cell and Molecular Biology Institute, College of Medicine, Soochow University, Suzhou 215123, China. tianangelt@163.com
authors.forename: Li-na
authors.initials: LN
authors.lastname: Tian
authors.forename: Xin
authors.initials: X
authors.lastname: Yu
authors.forename: Wan-rong
authors.initials: WR
authors.lastname: Bao
authors.forename: Yao
authors.initials: Y
authors.lastname: Chen
authors.forename: Ya-li
authors.initials: YL
authors.lastname: Han
authors.forename: Feng-juan
authors.initials: FJ
authors.lastname: Zhang
authors.forename: Jing-cheng
authors.initials: JC
authors.lastname: Miao
mesh_terms: D000256:Adenoviridae / Q000235:genetics; D018952:Antigens, CD34 / Q000276:immunology; D018929:Cell Culture Techniques / Q000379:methods; D049109:Cell Proliferation; D002469:Cell Separation / Q000379:methods; D002478:Cells, Cultured; D018920:Coculture Techniques / Q000379:methods; D005260:Female; D005312:Fetal Blood / Q000166:cytology* / Q000378:metabolism*; D018014:Gene Transfer Techniques / Q000295:instrumentation; D006412:Hematopoietic Stem Cells / Q000378:metabolism*; D006801:Humans; D018799:Intercellular Adhesion Molecule-1 / Q000378:metabolism; D053683:Oncostatin M / Q000235:genetics* / Q000378:metabolism*; D019718:Receptors, CXCR4 / Q000378:metabolism; D013234:Stem Cells / Q000378:metabolism*
chemical_list: D018952:Antigens, CD34; C508351:OSM protein, human; D019718:Receptors, CXCR4; D053683:Oncostatin M; D018799:Intercellular Adhesion Molecule-1