Advertisement

Maintenance of High Cytochrome P450 Expression in HepaRG Cell Spheroids in DMSO-Free Medium

  • A. V. AleksandrovaEmail author
  • O. A. Burmistrova
  • K. A. Fomicheva
  • D. A. Sakharov
Article

We studied the effects of DMSO and fibroblasts during HepaRG cell spheroid formation and conditions of their subsequent culturing on the levels of mRNA of the major cytochromes P450. A protocol of spheroid formation from differentiated HepaRG cells and their culturing in serum- and DMSO-free medium is developed.

Key Words

HepaRG differentiation cytochromes P450 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    S. Antherieu, C. Chesne, R. Li, S. Camus, A. Lahoz, L. Picazo, M. Turpeinen, A. Tolonen, J. Uusitalo, C. Guguen-Guillouzo, and A. Guillouzo, Stable expression, activity, and inducibility of cytochromes P450 in differentiated HepaRG cells. Drug Metab. Dispos., 38, No. 3, 516–525 (2010).CrossRefPubMedGoogle Scholar
  2. 2.
    V. V. Galatenko, M. Y. Shkurnikov, T. R. Samatov, A. V., Galatenko I. A. Mityakina, A. D. Kaprin, U. Schumacher, and A. G. Tonevitsky, Highly informative marker sets consisting of genes with low individual degree of differential expression. Sci. Rep., 5, 14967, doi:  10.1038/srep14967 (2015).
  3. 3.
    G. Langley, C. P. Austin, A. K. Balapure, L. S. Birnbaum, J. R. Bucher, J. Fentem, S. C. Fitzpatrick, J. R. Fowle, R. J. Kavlock, H. Kitano, B. A. Lidbury, A. R. Muotri, S. Q. Peng, D. Sakharov, T. Seidle, T. Trez, A. Tonevitsky, A. van de Stolpe, M. Whelan, and C. Willett, Lessons from toxicology: developing a 21st-century paradigm for medical research. Environ. Health Perspect, 123, No. 11, A268-A272 (2015).CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    E. L. LeCluyse, R. P. Witek, M. E. Andersen, and M. J. Powers, Organotypic liver culture models: meeting current challenges in toxicity testing. Crit. Rev. Toxicol., 42, No. 6, 501–548 (2012).CrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    D. V. Maltseva, N. A. Khaustova, N. N. Fedotov, E. O. Matveeva, A. E. Lebedev, M. U. Shkurnikov, V. V. Galatenko, U. Schumacher, and A. G. Tonevitsky, High-throughput identification of reference genes for research and clinical RT-qPCR analysis of breast cancer samples. J. Clin. Bioinforma, 3, No. 1, 13, doi:  10.1186/2043-9113-3-13 (2013).
  6. 6.
    E. M. Materne, A. P. Ramme, A. P. Terrasso, M. Serra, P. M. Alves, C. Brito, D. A. Sakharov, A. G. Tonevitsky, R. Lauster, and U. Marx, A multi-organ chip co-culture of neurospheres and liver equivalents for long-term substance testing. J. Biotechnol., 205, 36–46 (2015).CrossRefPubMedGoogle Scholar
  7. 7.
    E. M. Materne, A. G. Tonevitsky, and U. Marx, Chip-based liver equivalents for toxicity testing – organotypicalness versus cost-efficient high throughput. Lab. Chip, 13, No. 18, 3481–3495 (2013).CrossRefPubMedGoogle Scholar
  8. 8.
    M. Moisenovich, A. Tonevitsky, N. Maljuchenko, N. Kozlovskaya, I. Agapov, W. Volknandt, and J. Bereiter-Hahn, Endosomal ricin transport: involvement of Rab4- and Rab5-positive compartments. Histochem. Cell Biol., 121, No. 6, 429–439 (2004).CrossRefPubMedGoogle Scholar
  9. 9.
    B. Petzuch, N. Groll, M. Schwarz, and A. Braeuning, Application of HC-AFW1 hepatocarcinoma cells for mechanistic studies: regulation of cytochrome P450 2B6 expression by dimethyl sulfoxide and early growth response 1. Drug Metab. Dispos., 43, No. 11, 1727–1733 (2015).CrossRefPubMedGoogle Scholar
  10. 10.
    Y. Sakai, M. Koike, H. Hasegawa, K. Yamanouchi, A. Soyama, M. Takatsuki, T. Kuroki, K. Ohashi, T. Okano, and S. Eguchi, Rapid fabricating technique for multi-layered human hepatic cell sheets by forceful contraction of the fibroblast monolayer. PLoS One, 8, No. 7, doi:  10.1371/journal.pone.0070970 (2013).
  11. 11.
    T. R. Samatov, A. G. Tonevitsky, and U. Schumacher, Epithelialmesenchymal transition: focus on metastatic cascade, alternative splicing, non-coding RNAs and modulating compounds. Mol. Cancer, 12, No. 1, 107, doi:  10.1186/1476-4598-12-107 (2013).
  12. 12.
    Y. Takahashi, Y. Hori, T. Yamamoto, T. Urashima, Y. Ohara, and H. Tanaka, 3D spheroid cultures improve the metabolic gene expression profiles of HepaRG cells. Biosci. Rep., 35, No. 3, pii: e00208, doi:  10.1042/BSR20150034 (2015).
  13. 13.
    A. G. Tonevitsky, D. V. Maltseva, A. Abbasi, T. R. Samatov, D. A. Sakharov, M. U. Shkurnikov, A. E. Lebedev, V. V. Galatenko, A. I. Grigoriev, and H. Northoff, Dynamically regulated miRNA-mRNA networks revealed by exercise. BMC Physiol., 13, 9, doi: 10.1186/1472-6793-13-9 (2013).CrossRefPubMedPubMedCentralGoogle Scholar
  14. 14.
    C. C. Wang, S. Y. Lin, Y. H. Lai, Y. J. Liu, Y. L. Hsu, and J. J. Chen, Dimethyl sulfoxide promotes the multiple functions of the tumor suppressor HLJ1 through activator protein-1 activation in ells. PLoS One, 7, No. 4, doi:  10.1371/journal.pone.0033772 (2012).
  15. 15.
    Z. Wang, X. Luo, C. Anene-Nzelu, Y. Yu, X. Hong, N. H. Singh, L. Xia, S. Liu, and H. Yu, HepaRG culture in tethered spheroids as an in vitro three-dimensional model for drug safety screening. J. Appl. Toxicol., 35, No. 8, 909–917 (2015).CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • A. V. Aleksandrova
    • 1
    Email author
  • O. A. Burmistrova
    • 1
  • K. A. Fomicheva
    • 2
  • D. A. Sakharov
    • 1
    • 3
  1. 1.BioClinicum Research and Production CenterMoscowRussia
  2. 2.National Medical Research Radiology CenterMoscowRussia
  3. 3.Moscow State University of Mechanical Engineering (MAMI)MoscowRussia

Personalised recommendations