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Targeting early apoptotic genes in batch and fed‐batch CHO cell cultures
Author(s) -
Wong Danny Chee Furng,
Wong Kathy Tin Kam,
Nissom Peter Morin,
Heng Chew Kiat,
Yap Miranda Gek Sim
Publication year - 2006
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.20871
Subject(s) - apoptosis , gene , chinese hamster ovary cell , biology , cell culture , cell , computational biology , microbiology and biotechnology , chemistry , genetics
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 × 10 6 cells/mL) compared with the parental cell line (5 × 10 6 cells/mL). The recombinant interferon gamma (IFN‐γ) yields were increased by up to 2.5‐fold. Furthermore, it was observed that the IFN‐γ was more highly sialylated. © 2006 Wiley Periodicals, Inc.