Ex vivo organotypic culture system of precision-cut slices of human pancreatic ductal adenocarcinoma

Pancreatic ductal adenocarcinoma (PDAC) is one of the most overlooked cancers despite its dismal median survival time of 6 months. The biggest challenges in improving patient survival are late diagnosis due to lack of diagnostic markers, and limited treatment options due to almost complete therapy resistance. The past decades of research identified the dense stroma and the complex interplay/crosstalk between the cancer- and the different stromal cells as the main culprits for the slow progress in improving patient outcome. For better ex vivo simulation of this complex tumor microenvironment the models used in PDAC research likewise need to become more diverse. Depending on the focus of the investigation, several in vitro and in vivo models for PDAC have been established in the past years. Particularly, 3D cell culture such as spheroids and organoids have become more frequently used. This review aims to examine current PDAC in vitro models, their inherent limitations, and their successful implementations in research.

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Morphometric Assessment of Confocal Laser Endomicroscopy for Pancreatic Ductal Adenocarcinoma, an Ex-Vivo Pilot Study

Diagnostics ◽

10.3390/diagnostics10110923 ◽

2020 ◽

Vol 10 (11) ◽

pp. 923

Author(s):

Bogdan Silviu Ungureanu ◽

Daniel Pirici ◽

Simona Olimpia Dima ◽

Irinel Popescu ◽

Gheorghe Hundorfean ◽

...

Keyword(s):

Image Analysis ◽

Pilot Study ◽

Pancreatic Ductal Adenocarcinoma ◽

Ex Vivo ◽

Ductal Adenocarcinoma ◽

Confocal Laser Endomicroscopy ◽

Resection Margins ◽

Confocal Laser ◽

Cross Sectional ◽

Hole Area

Ex-vivo freshly surgical removed pancreatic ductal adenocarcinoma (PDAC) specimens were assessed using pCLE and then processed for paraffin embeding and histopathological diagnostic in an endeavour to find putative image analysis algorithms that might recognise adenocarcinoma. Methods: Twelve patients diagnosed with PDAC on endoscopic ultrasound and FNA confirmation underwent surgery. Removed samples were sprayed with acriflavine as contrast agent, underwent pCLE with an experimental probe and compared with previous recordings of normal pancreatic tissue. Subsequently, all samples were subjected to cross-sectional histopathology, including surgical resection margins for controls. pCLE records, as well as corespondant cytokeratin-targeted immunohistochemistry images were processed using the same morphological classifiers in the Image ProPlus AMS image analysis software. Specific morphometric classifiers were automatically generated on all images: Area, Hole Area (HA), Perimeter, Roundness, Integrated Optical Density (IOD), Fractal Dimension (FD), Ferret max (Fmax), Ferret mean (Fmean), Heterogeneity and Clumpiness. Results: After histopathological confirmation of adenocarcinoma areas, we have found that the same morphological classifiers could clearly differentiate between tumor and non-tumor areas on both pathology and correspondand pCLE (area, roundness, IOD, ferret and heterogeneity (p < 0.001), perimeter and hole area (p < 0.05). Conclusions: This pilot study proves that classical morphometrical classifiers can clearly differentiate adenocarcimoma on pCLE data, and the implementation in a live image-analysis algorithm might help in improving the specificity of pCLE in vivo diagnostic.

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The Role of Circular RNAs in Pancreatic Ductal Adenocarcinoma and Biliary-Tract Cancers

Cancers ◽

10.3390/cancers12113250 ◽

2020 ◽

Vol 12 (11) ◽

pp. 3250

Author(s):

Christopher Limb ◽

Daniel S. K. Liu ◽

Morten T. Veno ◽

Eleanor Rees ◽

Jonathan Krell ◽

...

Keyword(s):

Pancreatic Ductal Adenocarcinoma ◽

Biliary Tract ◽

Ex Vivo ◽

Expression Profiles ◽

Ductal Adenocarcinoma ◽

Circular Rnas ◽

Specific Expression ◽

Biliary Tract Cancers ◽

Cerna Network

Pancreatic Ductal Adenocarcinoma (PDAC) and biliary-tract cancers (BTC) often present at a late stage, and consequently patients have poor survival-outcomes. Circular RNAs (circRNAs) are non-coding RNA molecules whose role in tumourigenesis has recently been realised. They are stable, conserved and abundant, with tissue-specific expression profiles. Therefore, significant interest has arisen in their use as potential biomarkers for PDAC and BTC. High-throughput methods and more advanced bioinformatic techniques have enabled better profiling and progressed our understanding of how circRNAs may function in the competing endogenous RNA (ceRNA) network to influence the transcriptome in these cancers. Therefore, the aim of this systematic review was to describe the roles of circRNAs in PDAC and BTC, their potential as biomarkers, and their function in the wider ceRNA network in regulating microRNAs and the transcriptome. Medline, Embase, Scopus and PubMed were systematically reviewed to identify all the studies addressing circRNAs in PDAC and BTC. A total of 32 articles were included: 22 considering PDAC, 7 for Cholangiocarcinoma (CCA) and 3 for Gallbladder Cancer (GBC). There were no studies investigating Ampullary Cancer. Dysregulated circRNA expression was associated with features of malignancy in vitro, in vivo, and ex vivo. Overall, there have been very few PDAC and BTC tissues profiled for circRNA signatures. Therefore, whilst the current studies have demonstrated some of their functions in these cancers, further work is required to elucidate their potential role as cancer biomarkers in tissue, biofluids and biopsies.

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The Effects of 1α, 25-dihydroxyvitamin D3 and Transforming Growth Factor-β3 on Bone Development in an Ex Vivo Organotypic Culture System of Embryonic Chick Femora

PLoS ONE ◽

10.1371/journal.pone.0121653 ◽

2015 ◽

Vol 10 (4) ◽

pp. e0121653 ◽

Cited By ~ 6

Author(s):

Emma L. Smith ◽

Hassan Rashidi ◽

Janos M. Kanczler ◽

Kevin M. Shakesheff ◽

Richard O. C. Oreffo

Keyword(s):

Growth Factor ◽

Culture System ◽

Transforming Growth Factor ◽

Organotypic Culture ◽

Ex Vivo ◽

Bone Development ◽

Embryonic Chick ◽

Dihydroxyvitamin D3

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Pentraxin 3 is a stromally-derived biomarker for detection of pancreatic ductal adenocarcinoma

npj Precision Oncology ◽

10.1038/s41698-021-00192-1 ◽

2021 ◽

Vol 5 (1) ◽

Author(s):

Michelle R. Goulart ◽

Jennifer Watt ◽

Imran Siddiqui ◽

Rita T. Lawlor ◽

Ahmet Imrali ◽

...

Keyword(s):

Clinical Trial ◽

Pancreatic Ductal Adenocarcinoma ◽

Ex Vivo ◽

Pancreatic Stellate Cells ◽

Ductal Adenocarcinoma ◽

Diagnostic Biomarkers ◽

All Trans Retinoic Acid ◽

Desmoplastic Stroma ◽

Predictive Role

AbstractPancreatic ductal adenocarcinoma (PDAC), characterized by dense desmoplastic stroma laid down by pancreatic stellate cells (PSC), has no reliable diagnostic biomarkers for timely detection. A multi-center cohort of PDAC patients and controls (chronic pancreatitis, intra-ductal papillary neoplasms, gallstones and otherwise healthy) donated serum in an ethically approved manner. Serum PTX3 above 4.34 ng/mL has a higher sensitivity (86%, 95% confidence interval (CI): 65–97%) and specificity (86%, 95% CI: 79–91%), positive predictive value (97%) and likelihood ratio (6.05), and is superior when compared to serum CA19-9 and CEA for detection of PDAC. In vitro and ex vivo analyses of PTX3, in human PDAC samples, PSCs, cell lines and transgenic mouse model for PDAC, suggest that PTX3 originates from stromal cells, mainly PSC. In activated PSC, PTX3 secretion could be downregulated by rendering PSC quiescent using all-trans-retinoic acid (ATRA). PTX3 organizes hyaluronan in conjunction with tumor necrosis factor-stimulated gene 6 (TSG-6) and facilitates stellate and cancer cell invasion. In SCALOP clinical trial (ISRCTN96169987) testing chemo-radiotherapy without stromal targeting, PTX3 had no prognostic or predictive role. However, in STARPAC clinical trial (NCT03307148), stromal modulation by ATRA even at first dose is accompanied with serum PTX3 response in patients who later go on to demonstrate disease control but not those in whom the disease progresses. PTX3 is a putative stromally-derived biomarker for PDAC which warrants further testing in prospective, larger, multi-center cohorts and within clinical trials targeting stroma.

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Dynamic integration of enteric neural stem cells in ex vivo organotypic colon cultures

10.1101/2020.06.12.147652 ◽

2020 ◽

Author(s):

Georgina Navoly ◽

Conor J. McCann

Keyword(s):

Stem Cells ◽

Neural Stem Cells ◽

Culture System ◽

Organotypic Culture ◽

Ex Vivo ◽

Donor Cell ◽

Colonic Tissue ◽

Colonic Wall ◽

Donor Cells

AbstractEnteric neural stem cells (ENSC) have been identified as a possible treatment for enteric neuropathies. After in vivo transplantation, ENSC and their derivatives have been shown to engraft within colonic tissue, migrate and populate endogenous ganglia, and functionally integrate with the enteric nervous system. However, the mechanisms underlying the integration of donor ENSC, in recipient tissues, remains unclear. Here, using a modified ex vivo organotypic culture system we show that donor ENSC-derived cells integrate across the colonic wall in a dynamic fashion, across a three-week period. We further show that donor cells display two integrative patterns; longitudinal migration and medial invasion which allow donor cells to populate colonic tissue. Moreover, we demonstrate that significant remodelling of the intestinal ECM, and musculature, occurs upon transplantation to facilitate donor cell integration. Thus, our results provide critical evidence on the timescale, and mechanisms, which regulate donor ENSC integration within recipient gut tissue.

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