Viral Infection of Stem Cells

Dominik Haudenschild, from Jasper Yik 2009 UCDMC

• Obtain concentrated virus from GP2-293 packaging cells

• Tested on mmlv based pSuper siRNA and pMSCV vectors

• Seed bmsc at 7000 cells/cm² the night before infection

• Add polybrene to concentrated virus, final concentration of 10µg/ml

• Remove all media from stem cells

• Add 1ml concentrated virus with polybrene to cells on 6-well plate

• Leave cells in contact with virus no longer than 6 hours

• Remove virus, wash with PBS

• Apply fresh media, culture for 48 hours before applying selective agent (puromycin)

This yields ~80% infection for chondrocyte-like cell lines such as SW1353, and ~50% infection for primary stem cells