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Characterization of an in vitro cell culture bioreactor system to evaluate anti-neoplastic drug regimens

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A dynamic 3-dimensional tissue culture system has been developed that will allow for control of gemcitabine exposure to mimic concentration-time profiles measured from biologic samples. Gemcitabine was infused into a central reservoir. Media is mixed and delivered through hollow fiber capillaries, where it diffuses into the extracapillary space containing anchorage-dependent MDA-231 cells. To test for control of gemcitabine concentration-time profiles, drug was first infused through bioreactors without cells, and gemcitabine concentrations were measured with HPLC. Concentrations could be controlled to simulate 30-min and 2.5 h infusions, and were similar in both the lumen and extracapillary space. MDA-231 cells were then seeded into control (n = 4) and gemcitabine treatment (n = 4) groups, and maintained in culture for 2 weeks. Gemcitabine (5.3 mg) was infused over 30 min to the treatment group, and blank media to the control group. Accuracy of measured gemcitabine maximum concentration (C max) was 83.4%, and area under the curve (AUC), 106.2%, relative to pre-experimental theoretical values. With cells present, gemcitabine AUC in the extracapillary space was 32% of the value in the lumen. For the control group, 21.2 million cells (94.3% viable) were recovered, and for the gemcitabine-treated group, 16.8 million cells (87.1 % viable). Flow cytometry showed that 13.3 % of cells in the control group were in S-phase and 34.3 % in the gemcitabine-treated group were in S-phase (p = 0.003). In conclusion, gemcitabine concentration-time profiles could be accurately controlled through dosage, infusion rate, and pump flow rate, and cells could be recovered afterward to evaluate drug treatment.

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Acknowledgements

This work was supported in part by the Cancer Center Translational Breast Cancer Award to M.N.K.

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Authors and Affiliations

  1. Department of Experimental and Clinical Pharmacology, College of Pharmacy and Comprehensive Cancer Center, University of Minnesota, Minneapolis, USA

    Mark N. Kirstein

  2. Department of Experimental and Clinical Pharmacology, College of Pharmacy, University of Minnesota, Minneapolis, 55455, USA

    Richard C. Brundage & Dan E. Guire

  3. Department of Pharmaceutics, College of Pharmacy, University of Minnesota, Minneapolis, USA

    William F. Elmquist

  4. Department of Medicinal Chemistry, College of Pharmacy, University of Minnesota, Minneapolis, USA

    Rory P. Remmel

  5. Department of Medicine and Stem Cell Institute, University of Minnesota, Minneapolis, USA

    Paul H. Marker

  6. Department of Medicine, Department of Medicine and Cancer Center, University of Minnesota, Minneapolis, USA

    Douglas Yee

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  1. Mark N. Kirstein
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Address for offprints and correspondence: Mark N. Kirstein, Department of Experimental and Clinical Pharmacology, College of Pharmacy and Comprehensive Cancer Center, University of Minnesota, 308 Harvard St SE, Minneapolis, MN 55455, USA; Tel.: +1-612-624-5689; Fax: +1-612-625-3927

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Kirstein, M.N., Brundage, R.C., Elmquist, W.F. et al. Characterization of an in vitro cell culture bioreactor system to evaluate anti-neoplastic drug regimens. Breast Cancer Res Treat 96, 217–225 (2006). https://doi.org/10.1007/s10549-005-9004-z

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