Innervated mouse pancreas organoids as an ex vivo model to study pancreatic neuropathy in pancreatic cancer

H Erdinc Besikcioglu; Ümmügülsüm Yurteri; Enkhtsetseg Munkhbaatar; Linhan Ye; Fangfang Zhang; Alessandra Moretti; Carmen Mota Reyes; Candan Özoğul; Helmut Friess; Güralp O Ceyhan; Rouzanna Istvanffy; Ihsan Ekin Demir

doi:10.1016/j.xpro.2021.100935

Innervated mouse pancreas organoids as an ex vivo model to study pancreatic neuropathy in pancreatic cancer

STAR Protoc. 2021 Nov 17;2(4):100935. doi: 10.1016/j.xpro.2021.100935. eCollection 2021 Dec 17.

Authors

H Erdinc Besikcioglu^{1

2

3

4}, Ümmügülsüm Yurteri^{1

2

3}, Enkhtsetseg Munkhbaatar^{1

2

3}, Linhan Ye^{1

2

3

4}, Fangfang Zhang⁵, Alessandra Moretti⁵, Carmen Mota Reyes^{1

2

3}, Candan Özoğul⁶, Helmut Friess^{1

2

3}, Güralp O Ceyhan⁷, Rouzanna Istvanffy^{1

2

3}, Ihsan Ekin Demir^{1

2

3

7

8}

Affiliations

¹ Department of Surgery, Klinikum rechts der Isar, Technical University of Munich, School of Medicine, Munich, Germany.
² German Cancer Consortium (DKTK), Partner Site Munich, Munich, Germany.
³ CRC 1321 Modelling and Targeting Pancreatic Cancer, Munich, Germany.
⁴ Department of Histology and Embryology, Institution of Health Sciences, Gazi University, Ankara, Turkey.
⁵ Department of Internal Medicine I, Klinikum rechts der Isar, Technical University of Munich, School of Medicine, Munich, Germany.
⁶ Department of Histology and Embryology, Faculty of Medicine, University of Kyrenia, Kyrenia, Cyprus.
⁷ Department of General Surgery, HPB-Unit, School of Medicine, Acibadem Mehmet Ali Aydinlar University, Istanbul, Turkey.
⁸ Else Kröner Clinician Scientist Professor for Translational Pancreatic Surgery.

Abstract

Pancreatic cancer is characterized by bi-directional interactions between pancreatic cancer cells and stromal cells including neural cells. The absence of neural cells in pancreatic organoids limits the investigation of cell- cell interaction and tumor innervation. This protocol describes how to generate innervated wild type (WT) and Kras^+/LSLG12D Trp53^{fl/f l}p48^+/Cre (KPC) murine pancreatic organoids. To specifically investigate neurogenesis, organoids are co-cultured with iPSCs-derived neural crest cells, while co-culture with dorsal root ganglia explants is used for comparing organoids with mature neurons. For complete details on the use and execution of this protocol, please refer to Huch et al. (2013), Boj et al. (2015), and Demir et al. (2014).

Keywords: Cancer; Cell Differentiation; Cell culture; Cell isolation; Model Organisms; Neuroscience; Organoids; Stem Cells.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Cells, Cultured
Coculture Techniques / methods*
Mice
Models, Biological*
Organoids* / cytology
Organoids* / pathology
Pancreas / cytology*
Pancreatic Neoplasms / pathology*
Stromal Cells / cytology
Tumor Cells, Cultured / cytology