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Updated: Feb 16, 2026
Cell Co-culture Patterning Using Aqueous Two-phase Systems
Published on: March 26, 2013
Enrichment and Recovery of Mammalian Cells from Contaminated Cultures Using Aqueous Two-Phase Systems
Christopher J Luby1, Benjamin P Coughlin1, Charles R Mace1
1Department of Chemistry, Tufts University , 62 Talbot Avenue, Medford, Massachusetts 02155, United States.
Abstract:
This Article describes a density-based method for removing contaminants, including microorganisms and nonviable cells, from mammalian cell cultures using an aqueous two-phase system (ATPS). The properties of a 7% w/w polyethylene glycol (PEG)-11% w/w Ficoll ATPS can be tuned to prepare a biocompatible system that removes contaminants with little to no adverse effects on the viability or growth of the cultured cells after treatment. This system can be used to enrich cell culture populations for viable cells and to reduce the number of microorganism contaminants in a culture, which increases the chances of subsequent antibiotic treatments being successful. We test the effectiveness of our method in model contaminated cultures of both adherent (HeLa) and suspension (HL-60 II) mammalian cells contaminated with bacteria (E. coli) and yeast (S. cerevisiae). An average of 70.2 ± 4.6% of HeLa cells added to the system are subsequently recovered, and 55.9 ± 2.1% of HL-60 II cells are recovered. After sedimenting to the interface of the ATPS, these cells have an average viability of 98.0 ± 0.2% and 95.3 ± 2.2%, respectively. By removing unwanted cells, desired cell populations can be recovered, and cultures that would otherwise need to be discarded can continue to be used.
Insights
This study introduces a novel aqueous two-phase system (ATPS) for purifying mammalian cell cultures. The biocompatible method effectively removes contaminants like microorganisms and nonviable cells, preserving cell viability and enabling culture recovery.
Area of Science:
- Biotechnology
- Cell Biology
- Bioprocessing
Background:
- Mammalian cell cultures are susceptible to contamination by microorganisms and nonviable cells.
- Contamination can lead to culture loss and reduced efficacy of downstream processes.
- Effective methods for contaminant removal are crucial for maintaining cell culture integrity.
Purpose of the Study:
- To develop and evaluate a density-based method for removing contaminants from mammalian cell cultures.
- To assess the biocompatibility and effectiveness of an aqueous two-phase system (ATPS) for cell purification.
- To improve the recovery of viable cells and enhance the success of subsequent treatments.
Main Methods:
- Utilized a specific aqueous two-phase system (ATPS) composed of polyethylene glycol (PEG) and Ficoll.
- Tuned ATPS properties to create a biocompatible environment for cell purification.
- Tested the method on model contaminated cultures of adherent (HeLa) and suspension (HL-60 II) cells with bacterial (E. coli) and yeast (S. cerevisiae) contaminants.
Main Results:
- Achieved an average recovery of 70.2% for HeLa cells and 55.9% for HL-60 II cells.
- Recovered cells exhibited high viability (98.0% for HeLa, 95.3% for HL-60 II) after ATPS treatment.
- Demonstrated effective removal of bacterial and yeast contaminants from cell cultures.
Conclusions:
- The described ATPS method offers a biocompatible approach for removing contaminants from mammalian cell cultures.
- This technique can enrich viable cell populations and reduce microbial load, improving culture usability.
- The method presents a viable strategy to salvage contaminated cultures that might otherwise be discarded.
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