title: Tracing type 1 diabetic Tibet miniature pig's bone marrow mesenchymal stem cells in vitro by magnetic resonance imaging (1).
abstract: BACKGROUND
  Traditional cell-tracking methods fail to meet the needs of preclinical or clinical research. Thus, the aim of the present study was to establish a new method of double labeling bone marrow mesenchymal stem cells (BMSCs) from type 1 diabetic (T1D) minipigs with super-paramagnetic iron oxide (SPIO) and enhanced green fluorescent protein (eGFP) and tracing them using MRI in vitro.
  
  
  METHODS
  Isolated BMSCs from T1D minipigs were labeled with eGFP and different concentrations of SPIO. The effects of lentivirus (LV)-eGFP transfection and SPIO on the viability and growth curves of BMSCs were determined by Trypan blue exclusion, the 3-(4,5-dimethyl-2 thiazoyl)-2,5-diphenyl-2H-tetrazolium bromide assay and flow cytometry. Cellular ultrastructure was evaluated by transmission electron microscopy. Magnetic resonance imaging was used to evaluate BMSCs labeled with SPIO-eGFP complexes 6 weeks after labeling.
  
  
  RESULTS
  Expression of eGFP in BMSCs peaked 96 h after transfection with LV-eGFP. Prussian blue staining revealed scattered blue granules in the cytoplasm of SPIO-labeled cells. Transmission electron microscopy revealed that the dense granules aggregated mainly in secondary lysosomes. On MRI, T2* -weighted imaging was far more sensitive for SPIO-labeled BMSCs than other image sequences 3 and 6 weeks after the cells had been labeled with SPIO-eGFP.
  
  
  CONCLUSIONS
  We have developed a relatively simple and safe method for double labeling of BMSCs from T1D minipigs using SPIO and LV-eGFP and tracing them in vitro by MRI for 6 weeks.
authors: Department of Geriatrics, Qilu Hospital of Shandong University, Jinan; Department of Endocrinology, The 3rd Affiliated Hospital of Sun Yat-sen University.
authors: Kuan-Xiao
authors: KX
authors: Tang
authors: Jin-Hua
authors: JH
authors: Yan
authors: Yun-Feng
authors: YF
authors: Shen
authors: Bao-Ying
authors: BY
authors: Li
authors: Ying-Ming
authors: YM
authors: Chen
authors: Da-Yue
authors: DY
authors: Liu
authors: De-Dong
authors: DD
authors: Ma
authors: Jie
authors: J
authors: Li
authors: Hua
authors: H
authors: Liang
authors: Jian-Ping
authors: JP
authors: Weng
mesh_terms: D000818:Animals; D001854:Bone Marrow Cells / Q000000981:diagnostic imaging* / Q000378:metabolism / Q000648:ultrastructure; D049109:Cell Proliferation; D002470:Cell Survival; D058948:Cell Tracking / Q000379:methods; D002478:Cells, Cultured; D003922:Diabetes Mellitus, Type 1 / Q000097:blood*; D005290:Ferric Compounds / Q000737:chemistry; D049452:Green Fluorescent Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D008279:Magnetic Resonance Imaging / Q000379:methods*; D058185:Magnetite Nanoparticles / Q000737:chemistry; D059630:Mesenchymal Stem Cells / Q000000981:diagnostic imaging* / Q000378:metabolism / Q000648:ultrastructure; D046529:Microscopy, Electron, Transmission; D011859:Radiography; D015203:Reproducibility of Results; D013552:Swine; D013556:Swine, Miniature; D013997:Time Factors; D014162:Transfection
chemical_list: D005290:Ferric Compounds; D058185:Magnetite Nanoparticles; D049452:Green Fluorescent Proteins; C000499:ferric oxide
keywords: bone marrow mesenchymal stem cells; enhanced green fluorescent protein; magnetic resonance imaging; minipig; type 1 diabetes mellitus.; 骨髓间充质干细胞，增强型绿色荧光蛋白，磁共振成像，小型猪，1型糖尿病