Functional three-dimensional HepG2 aggregate cultures generated from an ultrasound trap: comparison with HepG2 spheroids.
Three-dimensional culture systems are an ideal in vitro model being capable of sustaining cell functionalities in a manner that resembles the in vivo conditions. In the present study, we developed an ultrasound trap-based technique to rapidly produce HepG2 hepatocarcinoma cell aggregates within 30 min. Enhanced junctional F-actin was observed at the points of cell-cell contact throughout the aggregates. HepG2 aggregates prepared by the ultrasound trap can be maintained in culture on a P-HEMA-coated surface for up to 3 weeks. The cells in these aggregates proliferated during the initial 3 days and cell number was consistent during the following maintenance period. Albumin secretion from these HepG2 aggregates recovered after 3 days of aggregate formation and remained relatively stable for the following 12 days. Cytochrome P450-1A1 activity was significantly enhanced after 6 days with maximal enzyme activity observed between 9 and 18 days. In addition, comparison experiments demonstrated that HepG2 aggregates generated by the ultrasound trap had comparable functional characterizations with HepG2 spheroids formed by a traditional gyrotatory-mediated method. Our results suggest that HepG2 aggregate cultures prepared through the ultrasound trap-based technique may provide a novel approach to produce in vitro models for hepatocyte functional studies.
Liu
Jian
J
Centre for Research in Biomedicine, Faculty of Applied Sciences, University of the West of England, Coldharbour Lane, Bristol BS16 1QY, UK. jian.liu@uwe.ac.uk
Kuznetsova
Larisa A
LA
Edwards
Gareth O
GO
Xu
Jinsheng
J
Ma
Mingwen
M
Purcell
Wendy M
WM
Jackson
Simon K
SK
Coakley
W Terence
WT
D000199:Actins / Q000378:metabolism; D000418:Albumins / Q000378:metabolism; D006528:Carcinoma, Hepatocellular / Q000473:pathology*; D002449:Cell Aggregation; D018929:Cell Culture Techniques; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D020099:Coated Materials, Biocompatible / Q000737:chemistry; D019363:Cytochrome P-450 CYP1A1 / Q000378:metabolism; D005782:Gels / Q000737:chemistry; D022781:Hepatocytes / Q000201:enzymology / Q000378:metabolism / Q000502:physiology*; D006801:Humans; D008113:Liver Neoplasms / Q000473:pathology*; D018874:Spheroids, Cellular / Q000473:pathology / Q000502:physiology*; D013997:Time Factors; D014465:Ultrasonics
D000199:Actins; D000418:Albumins; D020099:Coated Materials, Biocompatible; D005782:Gels; D019363:Cytochrome P-450 CYP1A1