Chapter 31
Drug Sensitivity Testing of Mycobacterium tuberculosis
Growing in a Hollow Fiber Bioreactor
A. Brett Mason and Ve ´ ronique Dartois
Abstract
Hollow fiber systems (HFSs) have been widely applied to study pharmacokinetic-pharmacodynamic
(PK-PD) relationships in antibiotic research and development. The system comprises a bundle of highdensity hollow capillary fibers that conduct a flow of medium with or without drug and an extra-capillary
space (ECS) inoculated with the pathogen of interest. The semipermeable membrane of the hollow fibers
allows for rapid exchange of small molecule drugs and solutes, while the pathogen is restricted to the ECS.
The unique properties of the HFS are (1) the ability to simulate any PK profile within the fibers and ECS,
including plasma or site-of-disease PK profiles, (2) the ability to simultaneously input several drugs with
different half-lives, (3) the ability to manipulate growth conditions such as medium composition, carbon
source, and pH, and (4) the ability to sample in both compartments in order to monitor drug concentrations and bacterial growth kinetics over time. The system is particularly suited for Mycobacterium tuberculosis research in a biosafety level 3 (BSL3) environment since pathogenic bacteria are sequestered in an
isolated compartment. The HFS was qualified by the European Medicines Agency for antituberculosis drug
development in 2015. Here, we describe the standard procedures used to study the growth kinetics of
M. tuberculosis in the HFS and the killing effect of first-line antituberculous drugs applied under simulated
human PK conditions. This animal-sparing and economical tool can be applied to optimize dosing
schedules that minimize emergence of resistance and to prioritize drug regimens that accelerate
sterilization.
Key words Tuberculosis, Bioreactor, Hollow fiber system (HFS), Pharmacokinetics (PK), Pharmacodynamics (PD), Central reservoir (CR), Extra-capillary space (ECS)
1 Introduction
Hollow fiber (HF) bioreactors are made of tubular capillary fibers
aligned lengthwise and enclosed within plastic cylinders. At each
terminus of the cylinder, the ends of the fibers are held in an
impermeable matrix “collar”. Fluid pumped from the central reservoir (CR, Fig. 1) enters through a terminal port on the cylinder
(Fig. 1) and flows inside the hollow fibers of the bioreactor.
Medium, small molecules and compounds can then perfuse freely
back and forth through the semipermeable fiber walls into the
Tanya Parish and Anuradha Kumar (eds.), Mycobacteria Protocols, Methods in Molecular Biology, vol. 2314,
https://doi.org/10.1007/978-1-0716-1460-0_31, © Springer Science+Business Media, LLC, part of Springer Nature 2021
715
Drug Sensitivity Testing of Mycobacterium tuberculosis
Growing in a Hollow Fiber Bioreactor
A. Brett Mason and Ve ´ ronique Dartois
Abstract
Hollow fiber systems (HFSs) have been widely applied to study pharmacokinetic-pharmacodynamic
(PK-PD) relationships in antibiotic research and development. The system comprises a bundle of highdensity hollow capillary fibers that conduct a flow of medium with or without drug and an extra-capillary
space (ECS) inoculated with the pathogen of interest. The semipermeable membrane of the hollow fibers
allows for rapid exchange of small molecule drugs and solutes, while the pathogen is restricted to the ECS.
The unique properties of the HFS are (1) the ability to simulate any PK profile within the fibers and ECS,
including plasma or site-of-disease PK profiles, (2) the ability to simultaneously input several drugs with
different half-lives, (3) the ability to manipulate growth conditions such as medium composition, carbon
source, and pH, and (4) the ability to sample in both compartments in order to monitor drug concentrations and bacterial growth kinetics over time. The system is particularly suited for Mycobacterium tuberculosis research in a biosafety level 3 (BSL3) environment since pathogenic bacteria are sequestered in an
isolated compartment. The HFS was qualified by the European Medicines Agency for antituberculosis drug
development in 2015. Here, we describe the standard procedures used to study the growth kinetics of
M. tuberculosis in the HFS and the killing effect of first-line antituberculous drugs applied under simulated
human PK conditions. This animal-sparing and economical tool can be applied to optimize dosing
schedules that minimize emergence of resistance and to prioritize drug regimens that accelerate
sterilization.
Key words Tuberculosis, Bioreactor, Hollow fiber system (HFS), Pharmacokinetics (PK), Pharmacodynamics (PD), Central reservoir (CR), Extra-capillary space (ECS)
1 Introduction
Hollow fiber (HF) bioreactors are made of tubular capillary fibers
aligned lengthwise and enclosed within plastic cylinders. At each
terminus of the cylinder, the ends of the fibers are held in an
impermeable matrix “collar”. Fluid pumped from the central reservoir (CR, Fig. 1) enters through a terminal port on the cylinder
(Fig. 1) and flows inside the hollow fibers of the bioreactor.
Medium, small molecules and compounds can then perfuse freely
back and forth through the semipermeable fiber walls into the
Tanya Parish and Anuradha Kumar (eds.), Mycobacteria Protocols, Methods in Molecular Biology, vol. 2314,
https://doi.org/10.1007/978-1-0716-1460-0_31, © Springer Science+Business Media, LLC, part of Springer Nature 2021
715