Effect of Molecular Weight of Phase Polymers on Partition of Cells in Aqueous Two-Phase Systems
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Effect of Molecular Weight of Phase Polymers on Partition of Cells in Aqueous Two-Phase Systems Ehsan Atefi1*, Ramila Joshi2, Hossein Tavana2* 1
Department of Mechanical Engineering, Manhattan College, Riverdale, NY 10471
2
Department of Biomedical Engineering, The University of Akron, Akron, OH 44325
*
Corresponding Authors
Ehsan Atefi, Ph.D. Mechanical Engineering Department Manhattan College, Riverdale, New York 10471 Tel.: 718-862-7756 Email: [email protected]
Hossein Tavana, Ph.D., P.Eng. Department of Biomedical Engineering The University of Akron, Akron, Ohio 44325 Tel.: 330-972-6031 Email: [email protected]
ABSTRACT Due to their aqueous environment and biocompatible polymers, aqueous two-phase systems (ATPS) provide a mild environment for partition of cells. A comprehensive understanding of cell partition in ATPS will facilitate cell separation and fractionation in ATPS for downstream cell-based analytical applications and various cellular and molecular biology studies. We report the effect of molecular weight of phase polymers on partition of cells between two aqueous phases and their interface. We generate three different ATPS by dissolving polyethylene glycol and dextran of different molecular weights in a standard cell culture medium of fixed composition. After suspending cells in each ATPS, we quantify the number of cells partitioned into each segregated phase and the interface between the two phases. Importantly, we use two-phase solutions of an identical interfacial tension from each ATPS to avoid the effect of interfacial tension on cell partition. Our results indicate that decreasing the molecular weight of one of the phase polymers results in distribution of a greater number of cells toward the phase rich in that polymer. Regardless of molecular weight of polymers used, two-phase solutions made with higher concentrations of polymers cause a significant shift toward cell partition to the interface. This study elucidates the role of polymer molecular weight on cell partition in ATPS and offers formulations for rapid and effective cell partition.
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INTRODUCTION Polymeric aqueous two-phase systems (ATPS) formed using polyethylene glycol (PEG) and dextran (DEX) offer a mild and highly aqueous environment to retain the functionality of biomolecules [1]. These systems have been extensively used to separate cell organelles, biomolecules, genetic materials, and cells [2-5]. Separation of large particles such as animal cells is exploited in biochemical and medical fields for analytical applications, gene analysis, screening of cell surface molecules, cell sorting, etc. [2,6,7]. Cell separation in ATPS has been utilized to develop novel three-dimensional cell printing and patterning techniques for disease modeling and tissue engineering applications. These inc
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