Exploiting the synergy between lysate and acoustic standing wave fields to increase the retroviral transduction of suspension cells

BIOT 3

Pascal R. Beauchesne, pbeauchesne@chml.ubc.ca1, Venkata S. Tayi, vtayi@chml.ubc.ca1, Bruce D. Bowen2, and James M. Piret, jpiret@chml.ubc.ca1. (1) Michael Smith Laboratories & Department of Chemical and Biological Engineering, University of British Columbia, 2185 East Mall, Vancouver, BC V6T 1Z4, Canada, (2) Department of Chemical and Biological Engineering, University of British Columbia, 2360 East Mall, Vancouver, BC V6T 1Z3, Canada
Recombinant retroviruses are effective therapeutic transgene delivery vehicles for stem cell-based gene therapy. However, the short half-life (~6 h) and low diffusivity (~6x10-8 cm2/s) of retroviruses often contribute to limited transduction efficiency. We have explored using whole cell lysate to improve the transduction efficiency of K-562 and TF-1 cells in MSCV-IRES-GFP virus-containing medium. Lysate addition increased transduction efficiency by 6-fold in t-flasks as well as in stirred suspension spinners and was a function of both lysate and target cell concentrations. When used in combination with a 1.95 MHz acoustic resonance filter, in which cells are retained within the pressure node planes while lysate and retroviral vectors are more readily displaced by fluid flow, transduction efficiency was increased by 2-fold more than for lysate alone leading to an overall increase of 12-fold. This approach utilizing the lysate-acoustic synergy can be readily scaled-up to transduce large numbers of cells.
 

Stem Cells: Quantifying, Modeling and Controlling Stem Cell Fate
8:20 AM-10:40 AM, Sunday, August 19, 2007 BCEC -- 108, Oral

Division of Biochemical Technology

The 234th ACS National Meeting, Boston, MA, August 19-23, 2007