title: Generation of an in vitro 3D PDAC stroma rich spheroid model.
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.
authors.affiliation: Department of Surgery, Baylor College of Medicine, Houston, TX, 77030, USA.
authors.forename: Matthew J
authors.initials: MJ
authors.lastname: Ware
authors.affiliation: Department of Surgery, Baylor College of Medicine, Houston, TX, 77030, USA.
authors.forename: Vazrik
authors.initials: V
authors.lastname: Keshishian
authors.affiliation: Department of Surgery, Baylor College of Medicine, Houston, TX, 77030, USA.
authors.forename: Justin J
authors.initials: JJ
authors.lastname: Law
authors.affiliation: Department of Surgery, Baylor College of Medicine, Houston, TX, 77030, USA.
authors.forename: Jason C
authors.initials: JC
authors.lastname: Ho
authors.affiliation: Department of Systems Medicine and Bio-engineering, Houston Methodist Research Institute, Houston, TX 77030, USA.
authors.forename: Carlos A
authors.initials: CA
authors.lastname: Favela
authors.affiliation: Department of Engineering, Swansea University, Swansea, United Kingdom.
authors.forename: Paul
authors.initials: P
authors.lastname: Rees
authors.affiliation: Pathology and Histology Core, Baylor College of Medicine, Houston, TX, 77030, USA.
authors.forename: Billie
authors.initials: B
authors.lastname: Smith
authors.affiliation: Pathology and Histology Core, Baylor College of Medicine, Houston, TX, 77030, USA.
authors.forename: Sayeeduddin
authors.initials: S
authors.lastname: Mohammad
authors.affiliation: Department of Surgical Oncology, University of Texas MD Anderson Cancer Center, Houston, TX 77030, USA.
authors.forename: Rosa F
authors.initials: RF
authors.lastname: Hwang
authors.affiliation: Department of Molecular and Cell Biology, Baylor College of Medicine, Houston, TX, 77030, USA.
authors.forename: Kimal
authors.initials: K
authors.lastname: Rajapakshe
authors.affiliation: Department of Molecular and Cell Biology, Baylor College of Medicine, Houston, TX, 77030, USA.
authors.forename: Cristian
authors.initials: C
authors.lastname: Coarfa
authors.affiliation: Department of Molecular and Cell Biology, Baylor College of Medicine, Houston, TX, 77030, USA.
authors.forename: Shixia
authors.initials: S
authors.lastname: Huang
authors.affiliation: Department of Molecular and Cell Biology, Baylor College of Medicine, Houston, TX, 77030, USA.
authors.forename: Dean P
authors.initials: DP
authors.lastname: Edwards
authors.affiliation: Department of Surgery, Baylor College of Medicine, Houston, TX, 77030, USA; Department of Chemistry, Rice University, Houston, TX 77005, USA; Department of Bioengineering, University of Houston, Houston, TX 77204, USA.
authors.forename: Stuart J
authors.initials: SJ
authors.lastname: Corr
authors.affiliation: Department of Nanomedicine, Houston Methodist Research Institute, Houston, TX 77030, USA. Electronic address: bianagodinv@gmail.com.
authors.forename: Biana
authors.initials: B
authors.lastname: Godin
authors.affiliation: Department of Surgery, Baylor College of Medicine, Houston, TX, 77030, USA.
authors.forename: Steven A
authors.initials: SA
authors.lastname: Curley
mesh_terms: D061249:Batch Cell Culture Techniques / Q000379:methods*; D021441:Carcinoma, Pancreatic Ductal / Q000473:pathology* / Q000503:physiopathology; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000502:physiology; D002470:Cell Survival / Q000502:physiology; D006801:Humans; D010190:Pancreatic Neoplasms / Q000473:pathology* / Q000503:physiopathology; D066330:Printing, Three-Dimensional; D018874:Spheroids, Cellular / Q000473:pathology*; D017154:Stromal Cells / Q000473:pathology; D023822:Tissue Engineering / Q000295:instrumentation / Q000379:methods*; D054457:Tissue Scaffolds
keywords: 3D tumor microenvironment; Human pancreatic stellate cells; Pancreatic cancer; Stroma