Transcriptional profiling of apoptotic pathways in batch and fed-batch CHO cell cultures

Chinese Hamster ovary (CHO) cells are regarded as one of the “work‐horses” for complex biotherapeutics production. In these processes, loss in culture viability occurs primarily via apoptosis, a genetically controlled form of cellular suicide. Using our “in‐house” developed CHO cDNA array and a mous...

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Published in:Biotechnology and bioengineering 2006-06, Vol.94 (2), p.373-382
Main Authors: Wong, Danny Chee Furng, Wong, Kathy Tin Kam, Lee, Yih Yean, Morin, Peter Nissom, Heng, Chew Kiat, Yap, Miranda Gek Sim
Format: Article
Language:English
Subjects:
Animals
Apoptosis
Apoptosis - genetics
Apoptosis - physiology
Apoptosis Regulatory Proteins - metabolism
Biological and medical sciences
Biotechnology
Cell culture
Cell Culture Techniques - methods
CHO Cells
Cricetinae
Culture Media
expression
fed batch
Fundamental and applied biological sciences. Psychology
Gene expression
Gene Expression Profiling
Gene Expression Regulation
microarray
Oligonucleotide Array Sequence Analysis
Rodents
Transcription, Genetic
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Summary:Chinese Hamster ovary (CHO) cells are regarded as one of the “work‐horses” for complex biotherapeutics production. In these processes, loss in culture viability occurs primarily via apoptosis, a genetically controlled form of cellular suicide. Using our “in‐house” developed CHO cDNA array and a mouse oligonucleotide array for time profile expression analysis of batch and fed‐batch CHO cell cultures, the genetic circuitry that regulates and executes apoptosis induction were examined. During periods of high viability, most pro‐apoptotic genes were down‐regulated but upon loss in viability, several early pro‐apoptotic signaling genes were up‐regulated. At later stages of viability loss, we detected late pro‐apoptotic effector genes such as caspases and DNases being up‐regulated. This sequential regulation of apoptotic genes showed that DNA microarrays could be used as a tool to study apoptosis. We found that in batch and fed‐batch cultures, apoptosis signaling occurred primarily via death receptor‐ and mitochondria‐mediated signaling pathways rather than endoplasmic reticulum‐mediated signaling. These insights provide a greater understanding of the regulatory circuitry of apoptosis during cell culture and allow for subsequent targeting of relevant apoptosis signaling genes to prolong cell culture. © 2006 Wiley Periodicals, Inc.
ISSN:0006-3592
1097-0290
DOI:10.1002/bit.20872