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Production of Retrovirus and Adenovirus Vectors for Gene Therapy: A Comparative Study Using Microcarrier and Stationary Cell Culture
Author(s) -
Wu SuhChin,
Huang Gloria YiaLing,
Liu JinHwang
Publication year - 2002
Publication title -
biotechnology progress
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 129
eISSN - 1520-6033
pISSN - 8756-7938
DOI - 10.1021/bp020026p
Subject(s) - microcarrier , retrovirus , viral vector , biology , virology , genetic enhancement , reporter gene , cell culture , vector (molecular biology) , transfection , gene delivery , microbiology and biotechnology , gene , virus , gene expression , genetics , recombinant dna
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.