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Optimization of adaptation parameters from adhesion cell culture in serum-containing media to suspension in chemically defined media by superlative box design

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Abstract

A new design of experiments—superlative box design (SBD), was adopted to optimize the adaptation of Chinese hamster ovary cells from adhesion culture to serum-free suspension culture. It is a general trend to use a serum-free medium instead of a serum-containing medium. The advantage of serum-free medium (chemically defended) is that it does not contain unknown components and avoids safety issues. SBD requires fewer experiments while ensuring a sufficient number of experiments and uniformity in the distribution of experiments amongst all the factors. Six factors were considered in this experimental design with 43 runs plus three more repeating center runs. The cell line was adapted to serum-free media by gradually reducing serum, and from adherent to suspension by rotating at various speeds in a shake flask. Response surface methodology was applied to find the optimum condition. The optimized cell density reached 7.02 × 105 cells/mL, calculated by the quadratic model.

Experiments validated the predicted cell adaptation with the maximum cell density. Three suspension runs were selected randomly to perform in the bioreactor to validate cell stability and production homogeneity. This study provides an efficient method to transfer adherent cells to suspension cells and is the first to successfully use SBD and establish a parameter quadratic optimization model.

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Abbreviations

CHO:

Chinese hamster ovary

DoE:

Design of Experiment

SBD:

Superlative box design

RSM:

Response surface methodology

CTLA-4:

Cytotoxic T-lymphocyte-associated antigen-4

DMEM:

Dulbecco’s modification of Eagle’s medium

FBS:

Fetal bovine serum

NMR:

Nuclear Magnetic Resonance

ELISA:

Enzyme-linked immunosorbent assay

ANOVA:

Analysis of variance

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Authors and Affiliations

  1. Department of Chemical Engineering, SUNY College of Environmental Science and Forestry, Syracuse, NY, 13210, USA

    Wanyue Cui & Shijie Liu

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  1. Wanyue Cui
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  2. Shijie Liu
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Contributions

WC conceived and designed research, conducted experiments, contributed analytical tools, analyzed data, and wrote the manuscript. SL revised the manuscript. All authors read and approved the manuscript.

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Correspondence to Shijie Liu.

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Appendix

Appendix

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Table 5 Six factor seven level SBD
Full size table

5.

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Cui, W., Liu, S. Optimization of adaptation parameters from adhesion cell culture in serum-containing media to suspension in chemically defined media by superlative box design. Cytotechnology 76, 39–52 (2024). https://doi.org/10.1007/s10616-023-00596-w

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