Autocrine stimulation of human hepatocytes triggers late DNA synthesis and stabilizes long-term differentiation in vitro

  • Authors:
    • Kerstin Leckel
    • Christoph Strey
    • Wolf O. Bechstein
    • Kim A. Boost
    • Marcus K.H. Auth
    • Amal El Makhfi
    • Eva Juengel
    • Steffen Wedel
    • Jon Jones
    • Dietger Jonas
    • Roman A. Blaheta
  • View Affiliations

  • Published online on: May 1, 2008     https://doi.org/10.3892/ijmm.21.5.611
  • Pages: 611-620
Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )
Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )


Abstract

Isolated human hepatocytes are of great value in investigating cell transplantation, liver physiology, pathology, and drug metabolism. Though hepatocytes possess a tremendous proliferative capacity in vivo, their ability to grow in culture is severely limited. We postulated that repeated medium change, common to most in vitro systems, may prevent long-term maintenance of hepato-specific functions and growth capacity. To verify our hypotheses we compared the DNA synthesis and differentiation status of isolated human hepatocytes, cultured in medium which was renewed every day or was not changed for 3 weeks (‘autocrine’ setting). Daily medium change led to rapid hepatocellular de-differentiation without any signs of DNA replication. In contrast, the autocrine setting allowed hepatocytes to become highly differentiated, demonstrated by an elevated ASGPr expression level, and increased albumin and fibrinogen synthesis and release. Cytokeratin 18 filaments were stably expressed, whereas cytokeratin 19 remained undetectable. Hepatocytes growing in an autocrine fashion were activated in the presence of hepatocyte growth factor (HGF), evidenced by c-Met phosphorylation. However, HGF response was not achieved when the culture medium was renewed daily. Furthermore, the autocrine setting evoked a late but strong interleukin 6 release into the culture supernatant, reaching maximum values after a 10-day cultivation period, and intense BrdU incorporation after a further 5-day period. Our data suggest that preservation of the same medium creates environmental conditions which allow hepatocytes to control their differentiation status and DNA synthesis in an autocrine fashion. Further studies are necessary to identify the key mediators involved in autocrine communication and to design the optimal culture configuration for clinical application.

Related Articles

Journal Cover

May 2008
Volume 21 Issue 5

Print ISSN: 1107-3756
Online ISSN:1791-244X

Sign up for eToc alerts

Recommend to Library

Copy and paste a formatted citation
x
Spandidos Publications style
Leckel K, Strey C, Bechstein WO, Boost KA, Auth MK, El Makhfi A, Juengel E, Wedel S, Jones J, Jonas D, Jonas D, et al: Autocrine stimulation of human hepatocytes triggers late DNA synthesis and stabilizes long-term differentiation in vitro. Int J Mol Med 21: 611-620, 2008.
APA
Leckel, K., Strey, C., Bechstein, W.O., Boost, K.A., Auth, M.K., El Makhfi, A. ... Blaheta, R.A. (2008). Autocrine stimulation of human hepatocytes triggers late DNA synthesis and stabilizes long-term differentiation in vitro. International Journal of Molecular Medicine, 21, 611-620. https://doi.org/10.3892/ijmm.21.5.611
MLA
Leckel, K., Strey, C., Bechstein, W. O., Boost, K. A., Auth, M. K., El Makhfi, A., Juengel, E., Wedel, S., Jones, J., Jonas, D., Blaheta, R. A."Autocrine stimulation of human hepatocytes triggers late DNA synthesis and stabilizes long-term differentiation in vitro". International Journal of Molecular Medicine 21.5 (2008): 611-620.
Chicago
Leckel, K., Strey, C., Bechstein, W. O., Boost, K. A., Auth, M. K., El Makhfi, A., Juengel, E., Wedel, S., Jones, J., Jonas, D., Blaheta, R. A."Autocrine stimulation of human hepatocytes triggers late DNA synthesis and stabilizes long-term differentiation in vitro". International Journal of Molecular Medicine 21, no. 5 (2008): 611-620. https://doi.org/10.3892/ijmm.21.5.611