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Comparative analysis of 3D culture methods on human HepG2 cells.
Luckert, Claudia; Schulz, Christina; Lehmann, Nadja; Thomas, Maria; Hofmann, Ute; Hammad, Seddik; Hengstler, Jan G; Braeuning, Albert; Lampen, Alfonso; Hessel, Stefanie.
Affiliation
  • Luckert C; Department of Food Safety, Federal Institute for Risk Assessment, Max-Dohrn-Straße 8-10, 10589, Berlin, Germany.
  • Schulz C; Department of Nutritional Toxicology, Institute of Nutritional Science, University of Potsdam, Arthur-Scheunert-Allee 114-116, 14558, Nuthetal, Germany.
  • Lehmann N; Department of Food Safety, Federal Institute for Risk Assessment, Max-Dohrn-Straße 8-10, 10589, Berlin, Germany.
  • Thomas M; Department of Food Safety, Federal Institute for Risk Assessment, Max-Dohrn-Straße 8-10, 10589, Berlin, Germany.
  • Hofmann U; Dr. Margarete Fischer-Bosch Institute of Clinical Pharmacology, Auerbachstraße 112, 70376, Stuttgart, Germany.
  • Hammad S; University of Tuebingen, Tuebingen, Germany.
  • Hengstler JG; Dr. Margarete Fischer-Bosch Institute of Clinical Pharmacology, Auerbachstraße 112, 70376, Stuttgart, Germany.
  • Braeuning A; University of Tuebingen, Tuebingen, Germany.
  • Lampen A; Leibniz Research Centre for Working Environment and Human Factors (IfADo), TU Dortmund University, Ardeystrasse 67, 44139, Dortmund, Germany.
  • Hessel S; Department of Forensic Medicine and Veterinary Toxicology, Faculty of Veterinary Medicine, South Valley University, 83523, Qena, Egypt.
Arch Toxicol ; 91(1): 393-406, 2017 Jan.
Article in En | MEDLINE | ID: mdl-26872951
ABSTRACT
Human primary hepatocytes represent a gold standard in in vitro liver research. Due to their low availability and high costs alternative liver cell models with comparable morphological and biochemical characteristics have come into focus. The human hepatocarcinoma cell line HepG2 is often used as a liver model for toxicity studies. However, under two-dimensional (2D) cultivation conditions the expression of xenobiotic-metabolizing enzymes and typical liver markers such as albumin is very low. Cultivation for 21 days in a three-dimensional (3D) Matrigel culture system has been reported to strongly increase the metabolic competence of HepG2 cells. In our present study we further compared HepG2 cell cultivation in three different 3D systems collagen, Matrigel and Alvetex culture. Cell morphology, albumin secretion, cytochrome P450 monooxygenase enzyme activities, as well as gene expression of xenobiotic-metabolizing and liver-specific enzymes were analyzed after 3, 7, 14, and 21 days of cultivation. Our results show that the previously reported increase of metabolic competence of HepG2 cells is not primarily the result of 3D culture but a consequence of the duration of cultivation. HepG2 cells grown for 21 days in 2D monolayer exhibit comparable biochemical characteristics, CYP activities and gene expression patterns as all 3D culture systems used in our study. However, CYP activities did not reach the level of HepaRG cells. In conclusion, the increase of metabolic competence of the hepatocarcinoma cell line HepG2 is not due to 3D cultivation but rather a result of prolonged cultivation time.
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Full text: 1 Collection: 01-internacional Database: MEDLINE Main subject: Gene Expression Regulation, Enzymologic / Toxicity Tests / Cell Culture Techniques / Hepatocytes / Tissue Scaffolds Type of study: Prognostic_studies Limits: Animals / Humans Language: En Journal: Arch Toxicol Year: 2017 Type: Article Affiliation country: Germany

Full text: 1 Collection: 01-internacional Database: MEDLINE Main subject: Gene Expression Regulation, Enzymologic / Toxicity Tests / Cell Culture Techniques / Hepatocytes / Tissue Scaffolds Type of study: Prognostic_studies Limits: Animals / Humans Language: En Journal: Arch Toxicol Year: 2017 Type: Article Affiliation country: Germany