Impact of cell growth morphology on retroviral transduction: Effect of contact inhibition

Authors
Citation
Yj. Kwon et Ca. Peng, Impact of cell growth morphology on retroviral transduction: Effect of contact inhibition, BIOTECH PR, 17(2), 2001, pp. 240-246
Citations number
25
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Biotecnology & Applied Microbiology",Microbiology
Journal title
BIOTECHNOLOGY PROGRESS
ISSN journal
8756-7938 → ACNP
Volume
17
Issue
2
Year of publication
2001
Pages
240 - 246
Database
ISI
SICI code
8756-7938(200103/04)17:2<240:IOCGMO>2.0.ZU;2-X
Abstract
Retrovirus-mediated gene transfer is one of the most commonly used methods to deliver, integrate, and express the gene of interest because the retrovi rus can insert the desired gene into the chromosome of the target cells wit h high stability. However, to deliver the gene successfully, the retrovirus requires active division to integrate reversely transcribed DNA into the c hromosome of target cells. In this study, we focused on the effect of cell- cell contact inhibition on the efficiency of retroviral transduction with t wo anchorage-dependent cell lines: NIH 3T3 and 293 cells. These two cell li nes have very different cell morphologies and growth patterns on surfaces. Human embryonic kidney epithelial 293 cells tend to stick together after di viding, while NIH 3T3 cells migrate to occupy available surface and spread. Experimental data indicate that the abatement of the transduction rate of 293 cells was initiated in the early stage of the culture, whereas effect o f contact inhibition of NIH 3T3 cells on the transduction rate became domin ating at the end of the culture period. Experimental results were also quan titatively illustrated by plotting normalized multiplicity of infection (MO I) versus normalized cell density. According to the outcomes, cell inoculat ion density plays an important role in optimizing the retroviral transducti on rate. The optimal time of retroviral transduction should be confined to the accelerating growth phase for 293 cells and at the exponential growth p hase for NIH 3T3 cells. The implication drawn from this study is that conta ct inhibition effect on retroviral transduction should be taken into accoun t for large-scale gene transfer systems such as the microcarrier bioreactor .