Determination of Drug Toxicity Using 3D Spheroids Constructed From an Immortal Human Hepatocyte Cell Line

S. J. Fey, Krzysztof Wrzesinski

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Abstract

Numerous publications have documented that the immortal cells
grown in three-dimensional (3D) cultures possess physiological
behavior, which is more reminiscent of their parental organ than
when the same cells are cultivated using classical two-dimensional
(2D) culture techniques. The goal of this study was to investigate
whether this observation could be extended to the determination of
LD50 values and whether 3D data could be correlated to in vivo
observations. We developed a noninvasive means to estimate the
amount of protein present in a 3D spheroid from it is planar area
(± 21%) so that a precise dose can be provided in a manner similar to
in vivo studies. This avoided correction of the actual dose given
based on a protein determination after treatment (when some cells
may have lysed). Conversion of published in vitro LC50 data (mM)
for six common drugs (acetaminophen, amiodarone, diclofenac,
metformin, phenformin, and valproic acid) to LD50 data (mg
compound/mg cellular protein) showed that the variation in LD50
values was generally less than that suggested by the original LC50
data. Toxicological analysis of these six compounds in 3D spheroid
culture (either published or presented here) demonstrated similar
LD50 values. Although in vitro 2D HepG2 data showed a poor
correlation, the primary hepatocyte and 3D spheroid data resulted
in a much higher degree of correlation with in vivo lethal blood
plasma levels. These results corroborate that 3D hepatocyte cultures
are significantly different from 2D cultures and are more
representative of the liver in vivo.
Original languageEnglish
JournalToxicological Sciences
Volume127
Issue number2
Pages (from-to)403-411
Number of pages9
ISSN1096-6080
DOIs
Publication statusPublished - 27. Mar 2012

Keywords

  • spheroid culture
  • Drug Toxicity
  • Planimetry
  • Protein to drug ratio
  • LD50 / LC50

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