Improving the time integral of viable cell concentration by overcoming cell death, namely apoptosis, is one of the widely used strategies for efficient production of therapeutic proteins. By establishing stable cell lines that overexpress anti-apoptotic genes or down-regulate pro-apoptotic genes, the final product yields can be enhanced as cells become more resistance to environmental stresses. From the selection of high-expressing clones to verification of anti-apoptotic activity, the method to construct a stable anti-apoptotic cell line is discussed in this chapter.
Keywords: Apoptosis; Cell culture; Chinese hamster ovary cells; Overexpression; Selection; Suspension adaptation; Therapeutic proteins; Transfection; Western blot analysis; siRNA.