title: [Enhanced green fluorescence protein reporter gene labeling of mesenchymal stem cells mediated by lentivirus].
abstract: OBJECTIVE
  To explore the effect of enhanced green fluorescence protein (EGFP) labeling mediated by lentivirus on the biophysical properties of mesenchymal stem cells (MSC), and whether the EGFP gene expression is permanent and stable.
  
  
  METHODS
  MSC were infected with EGFP lentivirus at different virus multiplicity of infection (MOI). EGFP positive rate was measured with fluorescent-activated cell scanning (FACS) analysis, and EGFP expression in MSC was investigated under a fluorescence microscope. Cell viability, proliferation, apoptosis and cell cycle were detected with trypan blue stain, MTT colorimetric assay, Hoechst stain and FACS analysis respectively. To evaluate the stability of EGFP expression, EGFP lentivirus infected MSC were harvested after cultured continuously in vitro for 2, 4, 8 or 16 weeks, and EGFP positive rate and fluorescence strength were detected with FACS analysis.
  
  
  RESULTS
  After infected with EGFP lentivirus (MOI = 20) for 96 h, EGFP positive rate of MSC was 97.39% +/- 0.68%. Cell viability, proliferation, apoptosis and cell cycle of MSC infected with EGFP lentivirus were unaffected, as compared with control MSC (P > 0.05). When cultured in vitro continuously for 2, 4, 8 or 16 weeks, EGFP positive rates of EGFP-MSC were 97.50% +/- 0.54%, 97.32% +/- 0.51%, 97.39% +/- 0.11%, and 97.48% +/- 0.13% respectively, while EGFP fluorescence strength were 440 +/- 13, 445 +/- 12, 458 +/- 13 and 456 +/- 16 respectively. Both EGFP positive rate and fluorescence strength kept in a stable level.
  
  
  CONCLUSIONS
  EGFP lentivirus can efficiently label MSC and has no significant effect on the biophysical properties of MSC. EGFP gene expression in MSC is permanent and stable. EGFP-MSC can be used for further cell tracing research.
authors.affiliation: Laboratory of Molecular Imaging, Department of Radiology, The Third Affiliated Hospital of Sun Yat-sen University, Guangzhou 510630, China.
authors.forename: Dan
authors.initials: D
authors.lastname: Li
authors.forename: Kang-shun
authors.initials: KS
authors.lastname: Zhu
authors.forename: Bin
authors.initials: B
authors.lastname: Zhou
authors.forename: Jin
authors.initials: J
authors.lastname: Wang
authors.forename: Rong-hua
authors.initials: RH
authors.lastname: Yan
authors.forename: Zheng-ran
authors.initials: ZR
authors.lastname: Li
authors.forename: Xiao-chun
authors.initials: XC
authors.lastname: Meng
authors.forename: Ming-sheng
authors.initials: MS
authors.lastname: Huang
authors.forename: Zai-bo
authors.initials: ZB
authors.lastname: Jiang
authors.forename: Hong
authors.initials: H
authors.lastname: Shan
mesh_terms: D000818:Animals; D015415:Biomarkers; D002478:Cells, Cultured; D017930:Genes, Reporter; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000235:genetics*; D016086:Lentivirus / Q000235:genetics*; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley
chemical_list: D015415:Biomarkers; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins