title: Lentivirus-mediated transfection of chondroitinase ABC gene without the bacterial leader sequence enables long-term secretion of functional chondroitinase ABC in human bone marrow stromal cells.
abstract: OBJECTIVE
  To test the feasibility of secretion of functional chondroitinase ABC (ChABC), a bacterial enzyme that promotes axonal regeneration after spinal cord injury, from human bone marrow stromal cells (hBMSCs).
  
  
  RESULTS
  A lentiviral-expression vector, Lenti6.3-ChABC-3F, carrying the ChABC-3F gene without the bacterial leader sequence (aa 1-24) was constructed. Transfection of these Lenti6.3-ChABC-3F lentivirus led to stable expression in and secrection of ChABC proteins from hBMSCs for at least ten passages in culture in vitro, which was demonstrated by QRT-PCR, immunostaining, western blotting and ELISA. Moreover, the secreted ChABC proteins exhibited similar functional activity as the commercially-available ChABC.
  
  
  CONCLUSIONS
  The lentivirus-mediated transfection of chondroitinase ABC gene without the bacterial leader sequence induced substantial long-term secretion of functional ChABC in hBMSCs.
authors.affiliation: Department of Neurology, Zhujiang Hospital, Southern Medical University, Guangzhou, China.
authors.forename: Yang
authors.initials: Y
authors.lastname: Guo
authors.affiliation: Department of Pediatrics, Robert H. Lurie Comprehensive Cancer Center, Stanley Manne Children's Research Institute, Northwestern University, Chicago, IL, USA.
authors.forename: Michael
authors.initials: M
authors.lastname: Klüppel
authors.affiliation: Department of Neurosurgery, National Key Clinic Department, Zhujiang Hospital, Southern Medical University, Guangzhou, China.
authors.forename: Hao
authors.initials: H
authors.lastname: Tang
authors.affiliation: Department of Neurology, Zhujiang Hospital, Southern Medical University, Guangzhou, China.
authors.forename: Sheng
authors.initials: S
authors.lastname: Tan
authors.affiliation: Department of Cardiovascular Medicine, Zhujiang Hospital, Southern, Medical University, Guangzhou, China.
authors.forename: Peidong
authors.initials: P
authors.lastname: Zhang
authors.affiliation: Department of Neurosurgery, National Key Clinic Department, Zhujiang Hospital, Southern Medical University, Guangzhou, China. czz1020@163.com.
authors.forename: Zhenzhou
authors.initials: Z
authors.lastname: Chen
mesh_terms: D002478:Cells, Cultured; D019765:Chondroitin ABC Lyase / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression; D020869:Gene Expression Profiling; D042822:Genomic Instability; D006801:Humans; D007118:Immunoassay; D016086:Lentivirus / Q000235:genetics; D059630:Mesenchymal Stem Cells / Q000378:metabolism*; D021382:Protein Sorting Signals / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism*; D014162:Transfection
chemical_list: D021382:Protein Sorting Signals; D011994:Recombinant Proteins; D019765:Chondroitin ABC Lyase
keywords: Bone marrow stromal cells; Chondroitin sulfate proteoglycans; Chondroitinase ABC; Lentivirus; Secretion