title: Efficient gene delivery into cell lines and stem cells using baculovirus.
abstract: Baculovirus is a promising vector for transducing numerous types of mammalian cells. We have developed hybrid baculovirus vectors and protocols for the efficient transduction of a variety of cell lines, primary cells and stem cells, including bone marrow-derived mesenchymal stem cells (BMSCs) and adipose-derived stem cells (ASCs). The hybrid vector enables intracellular minicircle formation and prolongs transgene expression. The advantages of this transduction protocol are that baculovirus supernatant alone needs to be added to cells growing in medium, and transduction occurs after only 4-6 h of incubation at room temperature (25 °C) with gentle shaking. The entire procedure, from virus generation to transduction, can be completed within 4 weeks. Compared with other transduction procedures, this protocol is simple and can confer efficiencies >95% for many cell types. This protocol has potential applications in tissue regeneration, as transduced cells continue to express transgenes after implantation. For example, transduction of rabbit ASCs (rASCs) with growth factor-encoding hybrid baculovirus vectors, as described as an example application in this protocol, enables robust and sustained growth factor expression, stimulates stem cell differentiation and augments tissue regeneration after implantation.
authors.affiliation: 1] Department of Chemical Engineering, National Tsing Hua University, Hsinchu, Taiwan. [2].
authors.forename: Li-Yu
authors.initials: LY
authors.lastname: Sung
authors.affiliation: 1] Department of Chemical Engineering, National Tsing Hua University, Hsinchu, Taiwan. [2].
authors.forename: Chiu-Ling
authors.initials: CL
authors.lastname: Chen
authors.affiliation: Department of Chemical Engineering, National Tsing Hua University, Hsinchu, Taiwan.
authors.forename: Shih-Yeh
authors.initials: SY
authors.lastname: Lin
authors.affiliation: Department of Chemical Engineering, National Tsing Hua University, Hsinchu, Taiwan.
authors.forename: Kuei-Chang
authors.initials: KC
authors.lastname: Li
authors.affiliation: Department of Chemical Engineering, National Tsing Hua University, Hsinchu, Taiwan.
authors.forename: Chia-Lin
authors.initials: CL
authors.lastname: Yeh
authors.affiliation: Department of Chemical Engineering, National Tsing Hua University, Hsinchu, Taiwan.
authors.forename: Guan-Yu
authors.initials: GY
authors.lastname: Chen
authors.affiliation: Department of Chemical Engineering, National Tsing Hua University, Hsinchu, Taiwan.
authors.forename: Chin-Yu
authors.initials: CY
authors.lastname: Lin
authors.affiliation: Department of Chemical Engineering, National Tsing Hua University, Hsinchu, Taiwan.
authors.forename: Yu-Chen
authors.initials: YC
authors.lastname: Hu
mesh_terms: D000818:Animals; D016367:Baculoviridae / Q000235:genetics*; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002460:Cell Line; D048091:Guided Tissue Regeneration; D008297:Male; D011817:Rabbits; D013234:Stem Cells / Q000166:cytology; D014161:Transduction, Genetic / Q000379:methods*