Production of Retrovirus and Adenovirus Vectors for Gene Therapy: A Comparative Study Using Microcarrier and Stationary Cell Culture

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• 作者：Suh-Chin Wu ; Gloria Yia-Ling Huang ; and Jin-Hwang Liu
• 刊名：Biotechnology Progress
• 出版年：2002
• 出版时间：June 2002
• 年：2002
• 卷：18
• 期：3
• 页码：617 - 622
• 全文大小：216K
• 年卷期：v.18,no.3(June 2002)
• ISSN：1520-6033

文摘

In gene therapy, retrovirus and adenovirus vectors are extensively used as gene-delivery vehicles and further large-scale processing of these viral vectors will beincreasingly important. This study examined stationary and microcarrier cell culturesystems with respect to the production of a retrovirus vector (encoding a monounithammerhead ribozyme gene with an intron) and an adenovirus vector (encoding areporter lacZ gene). Cytodex 1 and Cytodex 3 solid microcarriers were found to beable 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 growthbut produced significantly lower titers of retrovirus and adenovirus vector from theproducer cells. The specific retrovirus production was not proportionally related tothe specific growth rate of the producer cells. High MOI infection was essential forhigh-titer production of adenovirus vector in 293 cells. Hydrodynamic shear forces onmicrocarrier-grown cells increased the production yield for retrovirus vector butdecreased for adenovirus vector. The cellular productivity was much more efficientfor adenovirus vector produced in 293 cells as compared to the retrovirus vectorproduced in PA317-RCM1 cells. These findings can provide further insight into thefeasibility of applying microcarrier cell culture technology to produce gene-therapyvirus vectors.