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One of the main challenges in the development of bioprocesses based on cell transient expression is the commonly reported reduction of cell specific productivity at increasing cell densities. This is generally known as the cell density effect (CDE). Many efforts have been devoted to understanding the cell metabolic implications to this phenomenon in an attempt to design operational strategies to overcome it. A comprehensive analysis of the main studies regarding the CDE is provided in this work to better define the elements comprising its cause and impact. Then, examples of methodologies and approaches employed to achieve successful transient expression at high cell densities (HCD) are thoroughly reviewed. A critical assessment of the limitations of the reported studies in the understanding of the CDE is presented, covering the leading hypothesis of the molecular implications. The overall analysis of previous work on CDE may offer useful insights for further research into manufacturing of biologics. Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.


Jesús Lavado-García, Pol Pérez-Rubio, Laura Cervera, Francesc Gòdia. The cell density effect in animal cell-based bioprocessing: Questions, insights and perspectives. Biotechnology advances. 2022 Nov;60:108017

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PMID: 35809763

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