In this study we developed a PEEK-WC hollow fiber (HF) membrane bioreactor for the maintenance of human peripheral lymphocytes as model system for the in vitro investigation of disease pathogenesis, chemical effects and individual drug sensitivity. Peripheral lymphocytes isolated from donor's human buffy coat were cultured in the shell compartment of the PEEK-WC-HF bioreactor and stimulated with PHA 5microg/mL for the first 48h of culture to enhance cytokine production and cell proliferation. Thereafter, cells were cultured in the presence of Hypericum perforatum (St. John's wort) in order to induce cytochrome P450s enzymes, CYP2E, involved in the biotransformation of endogenous molecules and exogenous compounds. The metabolic activity of cells with respect to glucose consumption and oxygen uptake was maintained for all the culture time without the addition of mitogen. Two cytokines IL-2 and IL-10, which are specific pattern of lymphocytes T helper 1 and T helper 2, respectively, were produced in the bioreactor up to 14 days of culture. Lymphocytes were also able to biotransform acetaminophen through the formation of the main metabolite paracetamidofenil-beta-glucuronide, which is the product of glucuronidation reaction, as a result of the Hypericum perforatum administration that induced the catalytic activity of the CYP2E1. These results demonstrated the usefulness of the bioreactor as the support system that reproduces physiological parameters such as a constant perfusion of medium, nutrients and oxygen maintaining the in vitro integrity of lymphocyte viability and functions

Human lymphocyte PEEK-WC hollow fiber membrane bioreactor

CAROLEO, Maria Cristina;
2007

Abstract

In this study we developed a PEEK-WC hollow fiber (HF) membrane bioreactor for the maintenance of human peripheral lymphocytes as model system for the in vitro investigation of disease pathogenesis, chemical effects and individual drug sensitivity. Peripheral lymphocytes isolated from donor's human buffy coat were cultured in the shell compartment of the PEEK-WC-HF bioreactor and stimulated with PHA 5microg/mL for the first 48h of culture to enhance cytokine production and cell proliferation. Thereafter, cells were cultured in the presence of Hypericum perforatum (St. John's wort) in order to induce cytochrome P450s enzymes, CYP2E, involved in the biotransformation of endogenous molecules and exogenous compounds. The metabolic activity of cells with respect to glucose consumption and oxygen uptake was maintained for all the culture time without the addition of mitogen. Two cytokines IL-2 and IL-10, which are specific pattern of lymphocytes T helper 1 and T helper 2, respectively, were produced in the bioreactor up to 14 days of culture. Lymphocytes were also able to biotransform acetaminophen through the formation of the main metabolite paracetamidofenil-beta-glucuronide, which is the product of glucuronidation reaction, as a result of the Hypericum perforatum administration that induced the catalytic activity of the CYP2E1. These results demonstrated the usefulness of the bioreactor as the support system that reproduces physiological parameters such as a constant perfusion of medium, nutrients and oxygen maintaining the in vitro integrity of lymphocyte viability and functions
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Utilizza questo identificativo per citare o creare un link a questo documento: http://hdl.handle.net/20.500.11770/138882
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