Production of retrovirus and adenovirus vectors for gene therapy: a comparative study using microcarrier and stationary cell culture.
In gene therapy, retrovirus and adenovirus vectors are extensively used as gene-delivery vehicles and further large-scale processing of these viral vectors will be increasingly important. This study examined stationary and microcarrier cell culture systems with respect to the production of a retrovirus vector (encoding a monounit hammerhead ribozyme gene with an intron) and an adenovirus vector (encoding a reporter lacZ gene). Cytodex 1 and Cytodex 3 solid microcarriers were found to be able to provide good cell growth and high-titer vector production in suspension cultures. Porous microcarriers such as Cytopore 2 gave slightly lower but still efficient growth but produced significantly lower titers of retrovirus and adenovirus vector from the producer cells. The specific retrovirus production was not proportionally related to the specific growth rate of the producer cells. High MOI infection was essential for high-titer production of adenovirus vector in 293 cells. Hydrodynamic shear forces on microcarrier-grown cells increased the production yield for retrovirus vector but decreased for adenovirus vector. The cellular productivity was much more efficient for adenovirus vector produced in 293 cells as compared to the retrovirus vector produced in PA317-RCM1 cells. These findings can provide further insight into the feasibility of applying microcarrier cell culture technology to produce gene-therapy virus vectors.
Wu
Suh-Chin
SC
Department of Life Science, National Tsing Hua University, Hsinchu 30013, Taiwan. scwu@life.nthu.edu.tw
Huang
Gloria Yia-Ling
GY
Liu
Jin-Hwang
JH
D016475:3T3 Cells; D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D002455:Cell Division; D002460:Cell Line; D015316:Genetic Therapy; D005822:Genetic Vectors; D006801:Humans; D051379:Mice; D012190:Retroviridae / Q000235:genetics*