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Gene Transfer into Pluripotent Stem Cells via Lentiviral Transduction.
Naujok, Ortwin; Diekmann, Ulf; Elsner, Matthias.
Affiliation
  • Naujok O; Institute of Clinical Biochemistry, Hannover Medical School, 30625, Hannover, Germany. naujok.ortwin@mh-hannover.de.
  • Diekmann U; Institute of Clinical Biochemistry, Hannover Medical School, 30625, Hannover, Germany.
  • Elsner M; Institute of Clinical Biochemistry, Hannover Medical School, 30625, Hannover, Germany.
Methods Mol Biol ; 1341: 67-85, 2016.
Article in En | MEDLINE | ID: mdl-25762298
ABSTRACT
Recombinant lentiviral vectors are powerful tools to stably manipulate human pluripotent stem cells. They can be used to deliver ectopic genes, shRNAs, miRNAs, or any possible genetic DNA sequence into diving and nondividing cells. Here we describe a general protocol for the production of self-inactivating lentiviral vector particles and their purification to high titers by either ultracentrifugation or ultrafiltration. Next we provide a basic procedure to transduce human pluripotent stem cells and propagate clonal cell lines.
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Full text: 1 Collection: 01-internacional Database: MEDLINE Main subject: Transduction, Genetic / Gene Transfer Techniques / Lentivirus / Pluripotent Stem Cells / Genetic Vectors Limits: Humans Language: En Journal: Methods Mol Biol Journal subject: BIOLOGIA MOLECULAR Year: 2016 Type: Article Affiliation country: Germany

Full text: 1 Collection: 01-internacional Database: MEDLINE Main subject: Transduction, Genetic / Gene Transfer Techniques / Lentivirus / Pluripotent Stem Cells / Genetic Vectors Limits: Humans Language: En Journal: Methods Mol Biol Journal subject: BIOLOGIA MOLECULAR Year: 2016 Type: Article Affiliation country: Germany