Abstract
Pancreatic ductal adenocarcinoma (PDAC), the fourth leading cause of cancer death, has a 5-year survival rate of approximately 7–9%. The ineffectiveness of anti-PDAC therapies is believed to be due to the existence of a subpopulation of tumor cells known as cancer stem cells (CSCs), which are functionally plastic, and have exclusive tumorigenic, chemoresistant and metastatic capacities. Herein, we describe a 2D in vitro system for long-term enrichment of pancreatic CSCs that is amenable to biological and CSC-specific studies. By changing the carbon source from glucose to galactose in vitro, we force PDAC cells to utilize OXPHOS, resulting in enrichment of CSCs defined by increased CSC biomarker and pluripotency gene expression, greater tumorigenic potential, induced but reversible quiescence, increased OXPHOS activity, enhanced invasiveness, and upregulated immune evasion properties. This CSC enrichment method can facilitate the discovery of new CSC-specific hallmarks for future development into targets for PDAC-based therapies.
Highlights
Pancreatic ductal adenocarcinoma (PDAC), the fourth leading cause of cancer death, has a 5year survival rate of approximately 7–9%
Attempts to establish 2D cultures enriched in cancer stem cells (CSCs) using fluorescence-activated cell sorter (FACS) approaches have proven ineffective as marker-enriched CSCs quickly re-establish the heterogeneity of the culture present prior to sorting
We have developed a 2D in vitro system for long-term and sustained enrichment of pancreatic CSCs (PaCSCs) based on forced oxidative phosphorylation (OXPHOS)
Summary
Pancreatic ductal adenocarcinoma (PDAC), the fourth leading cause of cancer death, has a 5year survival rate of approximately 7–9%. Approaches that use specific signaling mechanisms to enrich and maintain CSC subpopulations with phenotypic plasticity in culture[5] could prove useful for (1) dissecting the underlying drivers of their plasticity and for (2) identifying new anti-CSC targets for drug development Toward this end, and based on our previous work demonstrating the metabolic differences that exist between PaCSCs and non-PaCSCs, we describe a longterm and sustained cell culture system enriched in PaCSCs based on forced oxidative phosphorylation (OXPHOS), defined by electron transport chain + ATP synthase. Based on our previous work demonstrating the metabolic differences that exist between PaCSCs and non-PaCSCs, we describe a longterm and sustained cell culture system enriched in PaCSCs based on forced oxidative phosphorylation (OXPHOS), defined by electron transport chain + ATP synthase In this culture system, cells increase the expression of PaCSC biomarkers and present better metabolic adaptation, plastic features such as a reversible quiescence-like state, and CSC-associated phenotypes, including chemoresistance, invasiveness, and immune evasive properties. This culture method sheds light on the link between previously unrecognized CSC features and mitochondrialdependent metabolism and represents a platform that could facilitate the discovery of CSC-specific properties that could lead to the development of new therapies against PaCSCs, which could improve the life expectancy of PDAC patients
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
