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Patient-derived, three-dimensional spheroid cultures provide a versatile translational model for the study of organ-confined prostate cancer

  • Johannes Linxweiler
  • Markus Hammer
  • Stefanie Muhs
  • Moritz Kohn
  • Alexej Pryalukhin
  • Christian Veith
  • Rainer M. Bohle
  • Michael Stöckle
  • Kerstin Junker
  • Matthias Saar
Original Article – Cancer Research

Abstract

Purpose

To generate and characterize 3D spheroid suspension cultures from radical prostatectomy (RP) specimens as a versatile model system for organ-confined prostate cancer (PCa).

Methods

Cancerous tissue samples from RP specimens were excised by a uropathologist. Preparation of 3D spheroids was done by mechanical disintegration and limited enzymatic digestion followed by serial filtration through 100 μm- and 40 μm-cell strainers. Thereafter, spheroids were cultured in a modified stem cell medium and characterized by a live/dead assay, whole-spheroid immunohistochemistry (IHC; CK5, CK8, AMACR, PSA, Ki67, AR, αSMA, Vimentin, E-Cadherin) and PSA-measurements in culture medium. Furthermore, their response to pharmaceutical treatment with docetaxel, bicalutamide, enzalutamide and abiraterone was tested.

Results

173 RP cases were included. The median preoperative PSA-level was 16.12 ng/ml [range 0.99;345], the median Gleason score was 7b [6;10]. 64 cases were excluded due to low tumor content in frozen sections (43) or to insufficient spheroid formation (21). In the remaining 109 cases, spheroids formed successfully and stayed viable for up to several months. IHC analysis revealed AR-, CK8-, and AMACR-positivity in nearly all cases, while CK5-positive cells were detectable only occasionally as were α-SMA and Vimentin. E-Cadherin was positive in most cases. Furthermore, spheroids proved to be amenable to cryopreservation. While abiraterone had no effect and docetaxel only a moderate effect, spheroid viability was markedly reduced upon bicalutamide and enzalutamide treatment.

Conclusions

Multicellular 3D spheroids can be generated from patient-derived RP tissue samples and serve as an innovative in vitro model of organ-confined PCa.

Keywords

3D In vitro model Organ-confined Organoid Prostate cancer Spheroid culture 

Notes

Acknowledgements

We are grateful to Maria Link, Helga Angeli, Gullan Hebel-Klebsch and Alexander Vogt for excellent technical assistance. We thank Andrea Hasenfus (Institut für Pathologie, St. Vincentius-Kliniken Karlsruhe) for her help during the initial phase of the project and Sebastian Hölters for his help with the immunohistochemistry and the cell culture assays. This work was supported by grants from HOMFOR (Homburger Forschungsförderung) to MSa.

Funding

This study was funded by a HOMFOR (Homburger Forschungsförderung) Grant to Matthias Saar.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflicts of interest to report.

Informed consent

Informed consent for the use of tissue samples from their radical prostatectomy specimens was obtained from all individual participants included in the study.

Supplementary material

432_2018_2803_MOESM1_ESM.pptx (2.7 mb)
Supplementary material 1 (PPTX 2766 KB)

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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Johannes Linxweiler
    • 1
  • Markus Hammer
    • 1
  • Stefanie Muhs
    • 1
  • Moritz Kohn
    • 1
  • Alexej Pryalukhin
    • 2
  • Christian Veith
    • 2
  • Rainer M. Bohle
    • 2
  • Michael Stöckle
    • 1
  • Kerstin Junker
    • 1
  • Matthias Saar
    • 1
  1. 1.Department of UrologySaarland UniversityHomburgGermany
  2. 2.Department of General and Surgical PathologySaarland UniversityHomburgGermany

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