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Preparation of Primary Acute Lymphoblastic Leukemia Cells in Different Cell Cycle Phases by Centrifugal Elutriation
Published on: November 10, 2017
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Near-Physiological Cell Cycle Synchronization with Countercurrent Centrifugal Elutriation
Johannes Möller1, Katrin Korte1, Ralf Pörtner1
1Institute of Bioprocess and Biosystems Engineering, Hamburg University of Technology, Hamburg, Germany.
Methods in Molecular Biology (Clifton, N.J.)
|December 21, 2019
Summary
Mammalian cell cultures exhibit heterogeneity affecting production. This study details using countercurrent centrifugal elutriation to isolate cell cycle-enriched subpopulations for studying metabolic regulations and productivities.
Area of Science:
- Biotechnology
- Cell Biology
- Bioprocess Engineering
Background:
- Mammalian cell cultures are often assumed homogeneous, but heterogeneities influence production rates.
- Variations in cell populations and cell cycle stages impact cultivation dynamics and productivity.
- CHO cell cultivations show metabolic regulations affecting productivity even in exponentially growing cultures.
Purpose of the Study:
- To investigate the influence of cell cycle heterogeneity on mammalian cell culture dynamics.
- To enable studies on cell cycle-related metabolic regulations and productivity variations.
- To present a method for generating cell cycle-enriched subpopulations without disrupting the culture.
Main Methods:
- Near-physiological countercurrent centrifugal elutriation for cell subpopulation enrichment.
- Detailed explanation of experimental setup and synchronization procedures.
- Overview of cell cycle, identified effects, and common methods.
Main Results:
- Countercurrent centrifugal elutriation effectively generates cell cycle-enriched subpopulations.
- The enrichment method minimizes interference with the investigated cell culture.
- Enables detailed study of cell cycle-specific metabolic regulations and productivities.
Conclusions:
- Cell cycle heterogeneity is a critical factor in mammalian cell culture productivity.
- Countercurrent centrifugal elutriation is a suitable method for isolating synchronized cell populations.
- This technique facilitates in-depth research into cell cycle-dependent bioprocess dynamics.
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