Gene Transfer into Pluripotent Stem Cells via Lentiviral Transduction

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.

Related Organizations

Hannover Medical School
Germany

Keywords

Pluripotent Stem Cells, Genetic Vectors, Lentivirus, Cell Culture Techniques, Gene Transfer Techniques, Ultrafiltration, HEK293 Cells, Transduction, Genetic, Humans, Ultracentrifugation, Cells, Cultured

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