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On-chip Cell Separator using Magnetic Bead-based Enrichment and Depletion of Various Cell Surface Markers

Estes, Matthew D.
http://rave.ohiolink.edu/etdc/view?acc_num=ucin1242661265

Abstract Details

2009, PhD, University of Cincinnati, Engineering : Biomedical Engineering.
The objective of this dissertation is the development of an innovative on-chip cell sorting system for multiple surface markers making use of both surface marker depletion and enrichment selection in series. The newly developed system uses magnetic separation technology to affect capture of cells in an ordered 2-dimensional matrix which presents a variety of benefits when compared with existing cell sorting techniques. A new on-chip cell sorting system has been proposed, designed, fabricated, and fully characterized for the separation of cells using multiple surface markers in this research work. The separation mechanism is based on the precise immobilization of magnetic beads coated with antibody for specific markers. The research presented here details the development of an electroplated paramagnetic microarray for the manipulation of magnetic beads using an externally supplied magnetic field, which shows better performances in separation when compared with previously reported work in paramagnetic array design, generating twice the local flux density gradient over a given area. The separation mechanism was tested and found to produce excellent magnetic bead capture at a variety of flow rates and bead concentrations, well in excess of 90 % efficiency. The paramagnetic array elements as well as microfluidic channels were constructed on a polymer wafer using cyclic olefin copolymer. The channel wafers and array wafers were bonded together to form a single biochip. The device was first tested with cell substitutes, using streptavidin coated magnetic beads paired with biotin coated cell substitutes, and anti-mouse IgG coated magnetic beads paired with mouse IgG coated cell substitutes. This was done with the goal of introducing a heterogeneous cell substitute population into the device, first removing the biotin labeled cell substitute subpopulation, and then capturing the mouse IgG labeled cell substitute subpopulation. Device performance was characterized as a function of both flow rate and incubation time. It was found that low flow rates and high incubation times produce the highest levels of specific cell capture. By first depleting for one marker and then enriching for a second marker a more specific subpopulation can be isolated than is possible with existing microscale magnetic bead-based cell sorters. The device presented in this work was applied to the subject of prostate cancer progression and tested using the LNCaP prostate cancer cell line. LNCaP cells were selectively labeled for either prostate specific membrane antigen or CD10. These two surface markers are increasingly identified as relating to the temporal succession of prostate cancer. The device proved capable of sorting for prostate cancer subpopulations labeled with either one or the other of these markers, which served as a model of a disease state where expression levels are transitioning. The microscale cell sorter proved capable of selective capture at a variety of flow rates and incubation times. The methodology detailed within this work adds a new device and method to on-chip cell separation techniques. The magnetic bead-based on-chip cell separator developed in this work can envisage a new on-chip cell separator in a low cost and disposable platform
Chong Ahn (Committee Chair)
William Ball (Committee Member)
Jeffrey Johnson (Committee Member)
Ian Papautsky (Committee Member)
Jason Shearn (Committee Member)
129 p.
Biomedical Research
magnetic bead-based cell sorting; MACS; Lab-on-a-Chip; microfluidics

Recommended Citations

Citations

  • Estes, M. D. (2009). On-chip Cell Separator using Magnetic Bead-based Enrichment and Depletion of Various Cell Surface Markers [Doctoral dissertation, University of Cincinnati]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=ucin1242661265

    APA Style (7th edition)

  • Estes, Matthew. On-chip Cell Separator using Magnetic Bead-based Enrichment and Depletion of Various Cell Surface Markers. 2009. University of Cincinnati, Doctoral dissertation. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=ucin1242661265.

    MLA Style (8th edition)

  • Estes, Matthew. "On-chip Cell Separator using Magnetic Bead-based Enrichment and Depletion of Various Cell Surface Markers." Doctoral dissertation, University of Cincinnati, 2009. http://rave.ohiolink.edu/etdc/view?acc_num=ucin1242661265

    Chicago Manual of Style (17th edition)

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© 2009, all rights reserved.
This open access ETD is published by University of Cincinnati and OhioLINK.