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

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Abstract

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.

Original languageEnglish (US)
Pages (from-to)217-225
Number of pages9
JournalBreast Cancer Research and Treatment
Volume96
Issue number3
DOIs
StatePublished - Apr 1 2006

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gemcitabine
Bioreactors
Cell Culture Techniques
Pharmaceutical Preparations
S Phase
Control Groups
Area Under Curve
In Vitro Techniques
Infusion Pumps

Keywords

  • Bioreactor
  • Gemcitabine
  • In vitro
  • MDA-231 cells
  • Pharmacokinetics
  • S-phase

Cite this

@article{c4ecd806e610400f91a8751dfc39146d,
title = "Characterization of an in vitro cell culture bioreactor system to evaluate anti-neoplastic drug regimens",
abstract = "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.",
keywords = "Bioreactor, Gemcitabine, In vitro, MDA-231 cells, Pharmacokinetics, S-phase",
author = "Kirstein, {Mark N.} and Brundage, {Richard C.} and Elmquist, {William F.} and Remmel, {Rory P.} and Marker, {Paul H.} and Guire, {Dan E.} and Douglas Yee",
year = "2006",
month = "4",
day = "1",
doi = "10.1007/s10549-005-9004-z",
language = "English (US)",
volume = "96",
pages = "217--225",
journal = "Breast Cancer Research and Treatment",
issn = "0167-6806",
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number = "3",

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TY - JOUR

T1 - Characterization of an in vitro cell culture bioreactor system to evaluate anti-neoplastic drug regimens

AU - Kirstein, Mark N.

AU - Brundage, Richard C.

AU - Elmquist, William F.

AU - Remmel, Rory P.

AU - Marker, Paul H.

AU - Guire, Dan E.

AU - Yee, Douglas

PY - 2006/4/1

Y1 - 2006/4/1

N2 - 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.

AB - 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.

KW - Bioreactor

KW - Gemcitabine

KW - In vitro

KW - MDA-231 cells

KW - Pharmacokinetics

KW - S-phase

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U2 - 10.1007/s10549-005-9004-z

DO - 10.1007/s10549-005-9004-z

M3 - Article

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AN - SCOPUS:33645748347

VL - 96

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EP - 225

JO - Breast Cancer Research and Treatment

JF - Breast Cancer Research and Treatment

SN - 0167-6806

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ER -