Rat hepatocyte spheroids formed by rocked technique maintain differentiated hepatocyte gene expression and function

Colleen M. Brophy, Jennifer L. Luebke-Wheeler, Bruce P. Amiot, Harris Khan, Rory P Remmel, Piero Rinaldo, Scott L. Nyberg

Research output: Contribution to journalArticle

104 Citations (Scopus)

Abstract

The culture of primary hepatocytes as spheroids creates an efficient three-dimensional tissue construct for hepatic studies in vitro. Spheroids possess structural polarity and functional bile canaliculi with normal differentiated function. Thus, hepatocyte spheroids have been proposed as the cell source in a variety of diagnostic, discovery, and therapeutic applications, such as a bioartificial liver. Using a novel rocking technique to induce spheroid formation, kinetics of spheroid formation, cell-cell adhesion, gene expression, and biochemical activities of rat hepatocyte spheroids were tested over 14 days of culture. Evidence was provided that the formation of spheroids occurred faster and with fewer nonadherent hepatocytes in rocked suspension culture compared to a traditional rotational system. Hepatocyte spheroids in rocked culture showed stable expression of more than 80% of 242 liver-related genes including those of albumin synthesis, urea cycle, phase I and II metabolic enzymes, and clotting factors. Biochemical activity of rocked spheroid hepatocytes was superior to monolayer culture of hepatocytes on tissue culture plastic and collagen. Conclusion: Spheroid formation by rocker technique was more rapid and more efficient than by rotational technique. Rocker-formed spheroids appear suitable for application in a bioartificial liver or as an in vitro liver tissue construct.

Original languageEnglish (US)
Pages (from-to)578-586
Number of pages9
JournalHepatology
Volume49
Issue number2
DOIs
StatePublished - Jun 24 2009

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Hepatocytes
Gene Expression
Artificial Liver
Liver
Bile Canaliculi
Blood Coagulation Factors
Cell Adhesion
Plastics
Urea
Albumins
Suspensions
Collagen
Enzymes
Genes

Cite this

Brophy, C. M., Luebke-Wheeler, J. L., Amiot, B. P., Khan, H., Remmel, R. P., Rinaldo, P., & Nyberg, S. L. (2009). Rat hepatocyte spheroids formed by rocked technique maintain differentiated hepatocyte gene expression and function. Hepatology, 49(2), 578-586. https://doi.org/10.1002/hep.22674

Rat hepatocyte spheroids formed by rocked technique maintain differentiated hepatocyte gene expression and function. / Brophy, Colleen M.; Luebke-Wheeler, Jennifer L.; Amiot, Bruce P.; Khan, Harris; Remmel, Rory P; Rinaldo, Piero; Nyberg, Scott L.

In: Hepatology, Vol. 49, No. 2, 24.06.2009, p. 578-586.

Research output: Contribution to journalArticle

Brophy, CM, Luebke-Wheeler, JL, Amiot, BP, Khan, H, Remmel, RP, Rinaldo, P & Nyberg, SL 2009, 'Rat hepatocyte spheroids formed by rocked technique maintain differentiated hepatocyte gene expression and function', Hepatology, vol. 49, no. 2, pp. 578-586. https://doi.org/10.1002/hep.22674
Brophy, Colleen M. ; Luebke-Wheeler, Jennifer L. ; Amiot, Bruce P. ; Khan, Harris ; Remmel, Rory P ; Rinaldo, Piero ; Nyberg, Scott L. / Rat hepatocyte spheroids formed by rocked technique maintain differentiated hepatocyte gene expression and function. In: Hepatology. 2009 ; Vol. 49, No. 2. pp. 578-586.
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