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Serum-free culture of primary human hepatocytes in a miniaturized hollow-fibre membrane bioreactor for pharmacological in vitro studies
Journal of tissue engineering and regenerative medicine, 2015-09, Vol.9 (9), p.1017-1026
Lübberstedt, Marc
Müller-Vieira, Ursula
Biemel, Klaus M.
Darnell, Malin
Hoffmann, Stefan A
Knöspel, Fanny
Wönne, Eva C
Knobeloch, Daniel
Nüssler, Andreas K
Gerlach, Jörg C
Andersson, Tommy B
Zeilinger, Katrin
2015
Volltextzugriff (PDF)
Details
Autor(en) / Beteiligte
Lübberstedt, Marc
Müller-Vieira, Ursula
Biemel, Klaus M.
Darnell, Malin
Hoffmann, Stefan A
Knöspel, Fanny
Wönne, Eva C
Knobeloch, Daniel
Nüssler, Andreas K
Gerlach, Jörg C
Andersson, Tommy B
Zeilinger, Katrin
Titel
Serum-free culture of primary human hepatocytes in a miniaturized hollow-fibre membrane bioreactor for pharmacological in vitro studies
Ist Teil von
Journal of tissue engineering and regenerative medicine, 2015-09, Vol.9 (9), p.1017-1026
Ort / Verlag
England: Blackwell Publishing Ltd
Erscheinungsjahr
2015
Quelle
MEDLINE
Beschreibungen/Notizen
Primary human hepatocytes represent an important cell source for in vitro investigation of hepatic drug metabolism and disposition. In this study, a multi‐compartment capillary membrane‐based bioreactor technology for three‐dimensional (3D) perfusion culture was further developed and miniaturized to a volume of less than 0.5 ml to reduce demand for cells. The miniaturized bioreactor was composed of two capillary layers, each made of alternately arranged oxygen and medium capillaries serving as a 3D culture for the cells. Metabolic activity and stability of primary human hepatocytes was studied in this bioreactor in the presence of 2.5% fetal calf serum (FCS) under serum‐free conditions over a culture period of 10 days. The miniaturized bioreactor showed functions comparable to previously reported data for larger variants. Glucose and lactate metabolism, urea production, albumin synthesis and release of intracellular enzymes (AST, ALT, GLDH) showed no significant differences between serum‐free and serum‐supplemented bioreactors. Activities of human‐relevant cytochrome P450 (CYP) isoenzymes (CYP1A2, CYP3A4/5, CYP2C9, CYP2D6, CYP2B6) analyzed by determination of product formation rates from selective probe substrates were also comparable in both groups. Gene expression analysis showed moderately higher expression in the majority of CYP enzymes, transport proteins and enzymes of Phase II metabolism in the serum‐free bioreactors compared to those maintained with FCS. In conclusion, the miniaturized bioreactor maintained stable function over the investigated period and thus provides a suitable system for pharmacological studies on primary human hepatocytes under defined serum‐free conditions. Copyright © 2012 John Wiley & Sons, Ltd.
Sprache
Englisch
Identifikatoren
ISSN: 1932-6254, 1932-7005
eISSN: 1932-7005
DOI: 10.1002/term.1652
Titel-ID: cdi_swepub_primary_oai_prod_swepub_kib_ki_se_132252603
Format
–
Schlagworte
bioreactor
,
Bioreactors
,
Cells, Cultured
,
Culture Media, Serum-Free
,
Cytochrome P-450 Enzyme System - metabolism
,
cytochrome P450
,
Drug Evaluation, Preclinical
,
Fluorescent Antibody Technique
,
Gene Expression Profiling
,
Hepatocytes - cytology
,
Hepatocytes - enzymology
,
Hepatocytes - metabolism
,
human hepatocytes
,
Humans
,
In Vitro Techniques
,
Medicin och hälsovetenskap
,
Membranes, Artificial
,
Miniaturization
,
perfusion
,
pharmacological studies
,
Regenerative medicine
,
serum-free
,
three-dimensional culture
,
Tissue engineering
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