Effect of cell lysates on retroviral transduction efficiency of cells in suspension culture.

Abstract:

:Recombinant retroviruses are effective vectors able to integrate transgenes into the target cell's genome to achieve longer-term expression. This study investigates the effect of cell lysis products, a common cell culture by-product, on the transduction of suspension cells by gammaretroviral vectors. Cell lysates derived from human and murine suspension cell lines significantly increased the transduction of human TF-1 and K-562 cell lines by gibbon ape leukemia virus-pseudotyped retroviral vectors without altering tropism. The transduction efficiency of TF-1 cells increased as a function of lysate concentration and decreased with increasing target cell concentrations. This was adequately predicted using a saturation equation based on the lysed-to-target cell concentration ratio, R, where: Fold increase = 1+Fold_(Max) (R/(K_(L)+R)). Lysate completely masked the effects of fibronectin when the two were added in combination. With protamine sulfate, the transduction efficiency was increased by lysate to 58% from 20% for protamine sulfate alone. Overall, the presence of cell lysate significantly influenced the outcome of the transduction process, either alone or in the presence of protamine sulfate or fibronectin.

journal_name

Biotechnol Bioeng

authors

Beauchesne PR,Bruce KJ,Bowen BD,Piret JM

doi

10.1002/bit.22634

subject

Has Abstract

pub_date

2010-04-15 00:00:00

pages

1168-77

issue

6

eissn

0006-3592

issn

1097-0290

journal_volume

105

pub_type

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