Abstract
BackgroundChinese hamster ovary cell line has been used routinely as a bioproduction factory of numerous biopharmaceuticals. So far, various engineering strategies have been recruited to improve the production efficiency of this cell line such as apoptosis engineering. Previously, it is reported that the caspase-7 deficiency in CHO cells reduces the cell proliferation rate. But the effect of this reduction on the CHO cell productivity remained unclear. Hence, in the study at hand the effect of caspase-7 deficiency was assessed on the cell growth, viability and protein expression. In addition, the enzymatic activity of caspase-3 was investigated in the absence of caspase-7.ResultsFindings showed that in the absence of caspase-7, both cell growth and cell viability were decreased. Cell cycle analysis illustrated that the CHO knockout (CHO-KO) cells experienced a cell cycle arrest in G2/M phase. This cell cycle arrest resulted in a 1.7-fold increase in the expression of luciferase in CHO-KO cells compared to parenteral cells. Furthermore, in the apoptotic situation the enzymatic activity of caspase-3 in CHO-KO cells was approximately 3 times more than CHO-K1 cells.ConclusionsThese findings represented that; however, caspase-7 deficiency reduces the cell proliferation rate but the resulted cell cycle arrest leads to the enhancement of recombinant protein expression. Moreover, increasing in the caspase-3 enzymatic activity compensates the absence of caspase-7 in the caspase cascade of apoptosis.
Highlights
Chinese hamster ovary cell line has been used routinely as a bioproduction factory of numerous biopharmaceuticals
In the study at hand, we aimed to identify the mechanisms by which caspase-7 affects cell growth, cell viability and cell cycle progression and investigate the effect of caspase-7 silencing on the expression yield of recombinant protein
Findings of this assay showed that caspase-3 activity had increased by up to threefold in Chinese hamster ovary (CHO)-KO cells undergoing apoptosis mediated by Sodium butyrate (NaBu) in comparison with CHO-K1 cells (Fig. 5)
Summary
Chinese hamster ovary cell line has been used routinely as a bioproduction factory of numerous biopharmaceuticals. CHO cell line is the most common mammalian expression system vastly used for the production of therapeutics [1, 2] This host cell is the cellular factory of about one third of all biopharmaceutical products approved by FDA since 1982 [3]. Caspase-7 has non-apoptotic roles, for instance in cell cycle progression To this end, it has been reported that the proteolytic activity of caspases affects the cell cycle proteins as their substrates [7]. In the study at hand, we aimed to identify the mechanisms by which caspase-7 affects cell growth, cell viability and cell cycle progression and investigate the effect of caspase-7 silencing on the expression yield of recombinant protein
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