title: [Screening and identification of mesenchymal stem cell strains to secret mouse interleukin-12 mediated with lenti-viral vector].
abstract: OBJECTIVE
  To screen the stable expression cell strains of mouse interleukin-12 (mIL-12) from mouse Mesenchymal Stem Cells (mMSCs) transfected with lenti-mIL-12 virus.
  
  
  METHODS
  The mIL-12 cDNA was amplified from plasmid pORF-mIL-12 (Invivogen) by PCR. The cDNA was subcloned into pENTR 11 to generate recombinant plasmid pENTR-mIL-12. Then, pENTR-mIL-12 was homologously recombinated with pLenti6/V5-Dest. The recombinant was named as pLenti6/V5-mIL-12 and confirmed by PCR and DNA sequencing. The Lenti6/V5-mIL-12 virus was packaged using 293FT cells. The Lenti-mIL-12-MSC monoclone was picked from the mMSCs infected by the Lenti6/V5-mIL-12 virus using Blasticidin and verified by RT-PCR and ELISA.
  
  
  RESULTS
  The recombinant pLenti6/V5-mIL-12 was constructed. The sequence of amplified mIL-12 gene was consistent with that reported in GenBank. By RT-PCR and ELISA, it was confirmed that the mIL-12 protein could be expressed and secreted into the supernatant of MSC strain culture.
  
  
  CONCLUSIONS
  The recombinant mMSC strains lentivirally engineered to secret mIL-12 were obtained.
authors.affiliation: State Key Laboratory of Biotherapy, West China Hospital, Sichuan University, Chengdu 610041, China.
authors.forename: Jian-Rong
authors.initials: JR
authors.lastname: Xu
authors.forename: Hong-Xia
authors.initials: HX
authors.lastname: Li
authors.forename: Guo-Qing
authors.initials: GQ
authors.lastname: Wang
authors.forename: Xiao-Bo
authors.initials: XB
authors.lastname: Du
authors.forename: Yu-Quan
authors.initials: YQ
authors.lastname: Wei
authors.forename: Ju-Mei
authors.initials: JM
authors.lastname: Zhao
mesh_terms: D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology*; D002460:Cell Line; D005260:Female; D005822:Genetic Vectors; D018664:Interleukin-12 / Q000096:biosynthesis* / Q000235:genetics; D016086:Lentivirus / Q000235:genetics* / Q000378:metabolism; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000378:metabolism*; D051379:Mice; D008807:Mice, Inbred BALB C; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D014162:Transfection
chemical_list: D011994:Recombinant Proteins; D018664:Interleukin-12