Human lymphocyte PEEK-WC hollow fiber membrane bioreactor.
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.
De Bartolo
Loredana
L
Institute on Membrane Technology, National Research Council of Italy, ITM-CNR, C/o University of Calabria, Cubo 17/C, Via P. Bucci, I-87030 Rende (CS), Italy. l.debartolo@itm.cnr.it
Piscioneri
Antonella
A
Cotroneo
Giuseppina
G
Salerno
Simona
S
Tasselli
Franco
F
Campana
Carla
C
Morelli
Sabrina
S
Rende
Maria
M
Caroleo
Maria Cristina
MC
Bossio
Marcello
M
Drioli
Enrico
E
D000082:Acetaminophen / Q000493:pharmacokinetics; D001577:Benzophenones; D001672:Biocompatible Materials; D019149:Bioreactors; D001709:Biotechnology; D001711:Biotransformation; D018929:Cell Culture Techniques; D005947:Glucose / Q000378:metabolism; D006801:Humans; D016753:Interleukin-10 / Q000096:biosynthesis; D007376:Interleukin-2 / Q000096:biosynthesis; D007659:Ketones; D008214:Lymphocytes / Q000166:cytology* / Q000276:immunology / Q000378:metabolism; D008567:Membranes, Artificial; D008855:Microscopy, Electron, Scanning; D011092:Polyethylene Glycols; D011108:Polymers
D001577:Benzophenones; D001672:Biocompatible Materials; C508609:IL10 protein, human; C508594:IL2 protein, human; D007376:Interleukin-2; D007659:Ketones; D008567:Membranes, Artificial; D011108:Polymers; D016753:Interleukin-10; C063834:polyetheretherketone; D000082:Acetaminophen; D011092:Polyethylene Glycols; D005947:Glucose