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Targeting early apoptotic genes in batch and fed-batch CHO cell cultures

✍ Scribed by Danny Chee Furng Wong; Kathy Tin Kam Wong; Peter Morin Nissom; Chew Kiat Heng; Miranda Gek Sim Yap

Publisher
John Wiley and Sons
Year
2006
Tongue
English
Weight
551 KB
Volume
95
Category
Article
ISSN
0006-3592

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✦ Synopsis

Based on the transcriptional profiling of CHO cell culture using microarray, four key early apoptosis signaling genes, Fadd, Faim, Alg-2, and Requiem, were identified and CHO GT (Gene Targeted) cell lines were developed by targeting these four genes. Two were CHO GT O cell lines overexpressing anti-apoptotic genes, Faim and Fadd DN and two were CHO GT KD cell lines involving knockdown of Alg-2 and Requiem which are pro-apoptotic genes using small interfering RNA (siRNA) technology. Comparisons of these CHO GT cell lines with the parental cell line in batch culture (BC) and fed-batch culture (FBC) were performed. Compared to parental cells, the CHO GT cell lines showed apoptosis resistance as they significantly delayed and/or suppressed initiator caspase-8 and -9 and executioner caspase-3 activities during culture. FBC of CHO GT cell lines reached significantly higher maximum viable cell densities (up to 9 Γ‚ 10 6 cells/mL) compared with the parental cell line (5 Γ‚ 10 6 cells/mL). The recombinant interferon gamma (IFN-g) yields were increased by up to 2.5-fold. Furthermore, it was observed that the IFN-g was more highly sialylated.

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