Development of decellularized matrices to produce an in vitro colorectal cancer model

Cancer has been responsible for the majority of deaths worldwide and for so has been considered the disease of the century. Namely, colorectal cancer (CRC) is one of most commonly diagnosed cancers, with 1.8 million new cases in 2018 (WHO data).

One of the reasons for the nonexistence of efficient therapies to tackle this disease is the lack of relevance of current models, which do not truly mimic the cancer environment. In fact, the environment that surrounds cancer cells is one of the main mediators of their behavior, namely for cell migration. A model capable to replicate cancer milieu would not only allow to reach a better understanding of the different hallmarks of cancer, but also, would allow to develop and test new therapies.

At the moment, Matrigel is the standard material used to mimic cancer environment. Nevertheless, this complex mixture of proteins secreted by mouse sarcoma cells has short self-life and lot-to-lot variability. Moreover, Matrigel is an undefined mixture, which hinders the determination of its influence on obtained results, namely which signals are influencing cell behavior [1].

Taking this in consideration, decellularized extracellular matrices (dECM) have emerged as promising material to emulate cancer microenvironment. Actually, dECM preserves specific tissues’ properties, such as composition, biomechanical properties and structure. To obtain the dECM, it is critical to implement an efficient method capable of removing all cellular components while maintaining minimal disruption of the ultrastructure and content [2].

In this study, we hypothesized that dECM produced by human adipose derived stem cells (hASCs) could be used to produce a more relying cancer microenvironment in vitro. For so, hASCs were cultured under αMEM supplemented with 10% FBS and 1% antibiotic/antimycotic for 6 weeks to produce high amounts of ECM. These matrices were decellularized using combination of techniques (physical, chemical and enzymatic) in order to better preserve the structure. The efficiency of the decellularization process was evaluated by DNA quantification and H&E staining for nucleus observation. Additionally, SEM analysis was used to observe the microstructure and the presence of collagen was investigated by Masson's trichrome staining. Finally, to investigate matrix composition SDS-PAGE and Western Blot were performed. Herein, we suggest that the production of dECM biomaterials can be used to develop in vitro cancer models capable of enhancing the outputs of cancer research, drug testing and personalized medicine.