A bioassay for the optimization of macrophage conditioned medium (MCM) as culture supplement used to promote hybridoma cell survival and growth

Authors: Hnasko, Robert; Lin, Alice; Stanker, Larry; McGarvey, Jeffery - Jeff

Submitted to: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 5/6/2018
Publication Date: 6/14/2018
Citation: Hnasko, R.M., Lin, A.V., Stanker, L.H., McGarvey, J.A. 2018. A bioassay for the optimization of macrophage conditioned medium (MCM) as culture supplement used to promote hybridoma cell survival and growth. Monoclonal Antibodies in Immunodiagnosis and Immunotherapy. 37(3):126-133. https://doi.org/10.1089/mab.2018.0008.
DOI: https://doi.org/10.1089/mab.2018.0008

Interpretive Summary: We developed a novel cellular bioassay that tests the bioactivity of macrophage conditioned medium (MCM), used as a cell culture supplement, in support of antibody producing cell lines. This is the first and only assay that allows for the optimization, validation and standardization of a MCM product that is widely used in cell culture systems to promote the survival and growth of hybridoma cells as part of monoclonal antibody selection and production.

Technical Abstract: Macrophage conditioned medium (MCM) is an important cell culture supplement used to support the survival and growth of newly fused hybridoma cells. The use of macrophage cells, as a part of hybridoma technology, has proven to be an effective and inexpensive source of growth factors that promote the early survival and growth of hybridoma cells. Despite the widespread use of MCM as a hybridoma culture supplement there is limited guidance and standardization for MCM production to achieve optimal hybridoma support. As an undefined supplement, significant variations in production of MCM may negatively impact hybridoma cell survival and growth. The lack of an available method for standardization of MCM bioactivity has limited validation, optimization, and commercial production. Consequently, variations in batch production of MCM may result in low quality MCM that limits hybridoma viability and negatively impacts monoclonal antibody production. In this report we describe a novel bioassay based on the newly generated MCM-dependent RMH359 hybridoma cell line that can be used to validate MCM bioactivity and standardize production. We demonstrate the utility of the RMH359 bioassay: (1) for evaluating MCM hybridoma bioactivity, (2) to define optimal conditions for production of MCM, and (3) as a method for MCM validation and standardization. In conclusion, the RMH359 cell bioassay provides a specific and sensitive assessment of MCM bioactivity in support of hybridoma cell survival and growth.