title: Efficient enhancement of lentiviral transduction efficiency in murine spermatogonial stem cells.
abstract: Spermatogonial stem cells (SSCs) are the foundation of spermatogenesis throughout postnatal life in male and have the ability to transmit genetic information to the subsequent generation. In this study, we have optimized the transduction efficiency of SSCs using a lentiviral vector by considering different multiplicity of infection (MOI), duration of infection, presence or absence of feeder layer and polycationic agents. We tested MOI of 5, 10 or 20 and infection duration of 6, 9 or 12 h respectively. After infection, cells were cultured for 1 week and as a result, the number of transduced SSCs increased significantly for MOI of 5 and 10 with 6 h of infection. When the same condition (MOI of 5 with 6 hours) was applied in presence or absence of STO feeder layer and infected SSCs were cultured for 3 weeks on the STO feeder layer, a significant increase in the number of transduced cells was observed for without the feeder layer during infection. We subsequently studied the effects of polycationic agents, polybrene and dioctadecylamidoglycyl spermine (DOGS), on the transduction efficiency. Compared with the polybrene treatment, the recovery rate of the transduced SSCs was significantly higher for the DOGS treatment. Therefore, our optimization study could contribute to the enhancement of germ-line modification of SSCs using lentiviral vectors and in generation of transgenic animals.
authors.affiliation: Department of Animal Science and Technology, Chung-Ang University, Ansung 456-756, Korea.
authors.forename: Bang-Jin
authors.initials: BJ
authors.lastname: Kim
authors.forename: Ki-Jung
authors.initials: KJ
authors.lastname: Kim
authors.forename: Yong-Hee
authors.initials: YH
authors.lastname: Kim
authors.forename: Yong-An
authors.initials: YA
authors.lastname: Lee
authors.forename: Byung-Gak
authors.initials: BG
authors.lastname: Kim
authors.forename: Chul Min
authors.initials: CM
authors.lastname: Cho
authors.forename: Hye-Ryeon
authors.initials: HR
authors.lastname: Kang
authors.forename: Chul Geun
authors.initials: CG
authors.lastname: Kim
authors.forename: Buom-Yong
authors.initials: BY
authors.lastname: Ryu
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D002478:Cells, Cultured; D005822:Genetic Vectors; D016086:Lentivirus / Q000235:genetics* / Q000378:metabolism; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D013091:Spermatogenesis / Q000235:genetics*; D013093:Spermatogonia / Q000166:cytology / Q000378:metabolism / Q000502:physiology*; D013234:Stem Cells / Q000378:metabolism / Q000502:physiology*; D014161:Transduction, Genetic / Q000379:methods