title: [Effects of lyophilized on the biological activities of lentiviral vector of bone morphogenetic protein 2].
abstract: OBJECTIVE
  To investigate the influence of lyophilization on the biological activity of recombinant lentiviral vectors of bone morphogenetic protein 2(BMP-2).
  
  
  METHODS
  Recombinant lenti-BMP-2 was constructed. lenti-BMP-2 was transfected with rat bone marrow stromal cells (BMSCs) by multiplicity infection (MOI) of 10, 25, 50, 100, 200. The infection efficiency was observed by X-gal staining. Under suitable conditions, the lenti-BMP-2 and 10% trehalose ratio of lyophilized protective agent was mixed into the lyophilization form. Before and after lyophilization, the effect of lenti-BMP-2 on the proliferation of BMSCs was evaluated by MTS assay. The expression of BMP-2 protein in the cells of lyophilized lenti-BMP-2 was detected by ELISA method. The expression of Runx-2, OCN, Col1 and OPN in BMSCs was detected by real-time PCR after transfection of lyophilized lenti-BMP-2. SPSS13.0 software package was used for statistical analysis.
  
  
  RESULTS
  X-gal staining showed an MOI of 100 pfu/cell, and stable transfection efficiency. Before and after lyophilization, no significant change was observed in regard to the effect of lyophilized lenti-BMP-2 on BMSCs proliferation (P>0.05). ELISA method showed that BMSCs transfected by lyophilized lenti-BMP-2 could express BMP-2 protein continuously and stably at a high level. Before and after lyophilization, the result of real-time PCR showed that no significant difference in the expression of OPN,Col1,OCN and Runx-2 in BMSCs (P>0.05).
  
  
  CONCLUSIONS
  Lyophilized lenti-BMP-2 with trehalose can maintain high activity for a long time as an effective and reliable storage method.
authors.affiliation: Shanghai Stomatological Disease Center, Shanghai Stomatological Disease Hospital. Shanghai 200001, China. E-mail:sammi510@126.com.
authors.forename: Xiao-Ling
authors.initials: XL
authors.lastname: Wei
authors.forename: Dao-Yang
authors.initials: DY
authors.lastname: Liu
authors.forename: Jie
authors.initials: J
authors.lastname: Pan
authors.forename: Pei-Jun
authors.initials: PJ
authors.lastname: Lu
authors.forename: Jun
authors.initials: J
authors.lastname: Zhao
mesh_terms: D000818:Animals; D001854:Bone Marrow Cells; D055396:Bone Morphogenetic Protein 2 / Q000235:genetics* / Q000378:metabolism; D002454:Cell Differentiation; D002478:Cells, Cultured; D005697:Galactosides; D005822:Genetic Vectors; D007211:Indoles; D059630:Mesenchymal Stem Cells; D010012:Osteogenesis; D051381:Rats; D060888:Real-Time Polymerase Chain Reaction; D011994:Recombinant Proteins; D014162:Transfection; D016212:Transforming Growth Factor beta
chemical_list: C525718:BMP2 protein, human; D055396:Bone Morphogenetic Protein 2; D005697:Galactosides; D007211:Indoles; D011994:Recombinant Proteins; D016212:Transforming Growth Factor beta; C499469:recombinant human bone morphogenetic protein-2; C044888:5-bromo-4-chloro-3-indolyl beta-galactoside