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Development of a diffusional microtitration device and a carbon fiber microsensor for potential drug influx/efflux studies on single cancer cell

Yi, Chen
http://rave.ohiolink.edu/etdc/view?acc_num=case1058284676

Abstract Details

1995, Doctor of Philosophy, Case Western Reserve University, Biomedical Engineering.
In this project we propose to test the hypothesis that anticancer drugs can be delivered into a single cell by a diffusional microburet (DMB), and the drug efflux, such as adriamycin efflux, can be monitored by microvoltammetry using a carbon fiber microelectrode. The combination of these two methods makes it possible to study the simultaneously influx, efflux and intracellular drug accumulation characteristics of resistant as well as sensitive cells as functions of time in a single cancer cell. The Diffusional Microburet has been invented and successfully realized to continuously deliver extremely small amounts of reagent/drug (in the order of femtomoles) into microscopic samples (in the order of femtoliters) or single biological cells by controlled diffusion. The delivery rate is typically equivalent to a hypothetical volumetric delivery rate of about 1 μL per year. It has been used to perform acid/base and complexometric microtitrations, and to deliver Lucifer yellow CH an inert fluorescent dye, into microscopic droplets and single cells. Adriamycin, an anticancer drug, is electrochemically detectable by a carbon fiber microelectrode. Adriamycin efflux from cancer cell populations was determined by using a carbon fiber microelectrode, in both drug sensitive and drug resistant cancer cell lines. A mathematical model was used to simulate the diffusion processes in the cell and the DMB, and the drug transport across the cell membrane in order to design the influx/efflux experiments on single cells. We found that the most promising experimental scheme to determine the efflux parameters is to measure the dynamic intracellular concentration and the influx from the DMB. The major advantages of the proposed experimental approach with respect to all existing techniques are: (1) Unlike other techniques which require a population of cells to work with, our proposed method makes it possible to detect drug efflux under simultaneous influx conditions on single cells. (2) No other technique can provide instantaneous and continuous influx and/or efflux data on single cancer cell. Our approach ensures a continuous reading with a seconds temporal resolution. While initial kinetics may play an important role in the onset of resistance mechanisms, only the techniques proposed here can provide data about that period with sufficient temporal resolution. (3) No other technique can enforce continuous drug uptake. A systematic study of influx rate dependence may lead to further clues as to how and why drug resistance develops and works. (4) In our proposed scheme, it is possible to deliver the drug directly into the cell without having to incubate it in a drug containing medium. Thus, the outside of the cell membrane need not be in contact with the drug for influx to occur. This circumstance may reveal currently unknown phenomena on initial intracellular drug distribution, free from any possible effects induced by the drug uptaking mechanism of the membrane. (5) We will be able to deliver H+, Ca2+, ATP, and other ions and/or drugs into the cancer cell while delivering Adriamycin, with the same DMB. The proposed technology thus allows for studying the coupling effects of ions and co-drug with the cancer drug
Miklos Gratzl (Advisor)
237 p.
Engineering, Biomedical
Microtitration device; Carbon fiber microsensor; Cancer cell

Recommended Citations

Citations

  • Yi, C. (1995). Development of a diffusional microtitration device and a carbon fiber microsensor for potential drug influx/efflux studies on single cancer cell [Doctoral dissertation, Case Western Reserve University]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=case1058284676

    APA Style (7th edition)

  • Yi, Chen. Development of a diffusional microtitration device and a carbon fiber microsensor for potential drug influx/efflux studies on single cancer cell. 1995. Case Western Reserve University, Doctoral dissertation. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=case1058284676.

    MLA Style (8th edition)

  • Yi, Chen. "Development of a diffusional microtitration device and a carbon fiber microsensor for potential drug influx/efflux studies on single cancer cell." Doctoral dissertation, Case Western Reserve University, 1995. http://rave.ohiolink.edu/etdc/view?acc_num=case1058284676

    Chicago Manual of Style (17th edition)

case1058284676
501
© 1995, all rights reserved.
This open access ETD is published by Case Western Reserve University School of Graduate Studies and OhioLINK.