Generation of an in vitro 3D PDAC stroma rich spheroid model

Matthew J. Ware, Vazrik Keshishian, Justin J. Law, Jason C. Ho, Carlos A. Favela, Paul Rees, Billie Smith, Sayeeduddin Mohammad, Rosa F. Hwang, Kimal Rajapakshe, Cristian Coarfa, Shixia Huang, Dean P. Edwards, Stuart J. Corr, Biana Godin, Steven A. Curley

Research output: Contribution to journalArticlepeer-review

70 Scopus citations

Abstract

Pancreatic ductal adenocarcinoma (PDAC) is characterized by a prominent desmoplastic/stromal reaction, which contributes to the poor clinical outcome of this disease. Therefore, greater understanding of the stroma development and tumor-stroma interactions is highly required. Pancreatic stellate cells (PSC) are myofibroblast-like cells located in exocrine areas of the pancreas, which as a result of inflammation produced by PDAC migrate and accumulate in the tumor mass, secreting extracellular matrix components and producing the dense PDAC stroma. Currently, only a few orthotopic or ectopic animal tumor models, where PDAC cells are injected into the pancreas or subcutaneous tissue layer, or genetically engineered animals offer tumors that encompass some stromal component. Herein, we report generation of a simple 3D PDAC in vitro micro-tumor model without an addition of external extracellular matrix, which encompasses a rich, dense and active stromal compartment. We have achieved this in vitro model by incorporating PSCs into 3D PDAC cell culture using a modified hanging drop method. It is now known that PSCs are the principal source of fibrosis in the stroma and interact closely with cancer cells to create a tumor facilitatory environment that stimulates local and distant tumor growth. The 3D micro-stroma models are highly reproducible with excellent uniformity, which can be used for PDAC-stroma interaction analysis and high throughput automated drug-screening assays. Additionally, the increased expression of collagenous regions means that molecular based perfusion and cytostaticity of gemcitabine is decreased in our Pancreatic adenocarcinoma stroma spheroids (PDAC-SS) model when compared to spheroids grown without PSCs. We believe this model will allow an improved knowledge of PDAC biology and has the potential to provide an insight into pathways that may be therapeutically targeted to inhibit PSC activation, thereby inhibiting the development of fibrosis in PDAC and interrupting PSC-PDAC cell interactions so as to inhibit cancer progression.

Original languageEnglish (US)
Pages (from-to)129-142
Number of pages14
JournalBiomaterials
Volume108
DOIs
StatePublished - Nov 1 2016
Externally publishedYes

Bibliographical note

Funding Information:
B.G. acknowledges the financial support from NIH U54CA143837 Physical Sciences and Oncology grant, NIH 1U54CA151668-01 Cancer Center for Nanotechnology Excellence grant and 1R21HD082947 . MJW, SC and SJC acknowledge the financial support from Kanzius Cancer Research Foundation . The authors would also like to acknowledge kind assistance of Kemi Cui (HMRI Advanced Cellular and Tissue Microscope Core Facility) for his help with acquiring color histological images. SH, CC and KR acknowledges the financial support from Cancer Prevention & Research Institute of Texas Proteomics & Metabolomics Core Facility Support Award (RP120092) (DPE and SH) and NCI Cancer Center Support Grant to Antibody-based Proteomics Core/Shared Resource (P30CA125123) (DPE and SH). The metabolomics core is also supported by CPRIT Core facility award RP120092- DPE is PI). The authors would also like to acknowledge the LC-MS/MS work performed by Sri Ramya Donepudi and Nagireddy Putluri in Advanced Technology Core, Baylor College of Medicine. The Advanced Technology Core is supported by the following grants: NCI/2P30CA125123-09 Shared Resources Metabolomics core and funds from Dan L. Duncan Cancer Center (DLDCC), Baylor college of Medicine. Appendix A

Publisher Copyright:
© 2016 Elsevier Ltd

Keywords

  • 3D tumor microenvironment
  • Human pancreatic stellate cells
  • Pancreatic cancer
  • Stroma

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