Towards unravelling the kinetics of an acute myeloid leukaemia model system under oxidative and starvation stress: a comparison between two- and three-dimensional cultures

Abstract

A great challenge when conducting ex vivo studies of leukaemia is the construction of an appropriate experimental platform that would recapitulate the bone marrow (BM) environment. Such a 3D scaffold system has been previously developed in our group [1]. Additionally to the BM architectural characteristics, parameters such as oxygen and glucose concentration are crucial as their value could differ between patients as well as within the same patient at different stages of treatment, consequently affecting the resistance of leukaemia to chemotherapy. The effect of oxidative and glucose stress-at levels close to human physiologic ones-on the proliferation and metabolic evolution of an AML model system (K-562 cell line) in conventional 2D cultures as well as in 3D scaffolds were studied. We observed that the K-562 cell line can proliferate and remain alive for 2 weeks in medium with glucose close to physiological levels both in 20 and 5% O2. We report interesting differences on the cellular response to the environmental, i.e., oxidative and/or nutritional stress stimuli in 2D and 3D. Higher adaptation to oxidative stress under non-starving conditions is observed in the 3D system. The glucose level in the medium has more impact on the cellular proliferation in the 3D compared to the 2D system. These differences can be of significant importance both when applying chemotherapy in vitro and also when constructing mathematical tools for optimisation of disease treatment.

Chemoradiotherapy screening in a novel biomimetic polymer based pancreatic cancer model

“…The scaffolds were fabricated via the thermal induced phase separation method prepared as reported previously. 7,[62][63][64] Briey, PU beads (Noveon, Belgium) were dissolved in dioxane (5% w/v) (99.8% anhydrous pure, Sigma-Aldrich, UK) followed by quenching at À80 C for 3 h. The solvent was then removed by freeze drying in a poly-ethylene glycol (PEG) bath at À15 C under 0.01 mbar vacuum pressure. The scaffolds were then cut into 5 Â 5 Â 5 mm 3 cubes and sterilised by exposing them to 70% ethanol (3 h) and UV ray (1 h).…”

“…As mentioned previously, Gemcitabine (GEM), a standard chemotherapeutic agent against PDAC [73][74][75]90,91 was introduced to the culture medium surrounding the scaffolds at week 4 of culture and at different concentrations, i.e. 10, 50 and 100 mM, for 1 feeding cycle, i.e.…”

A 3D bioinspired highly porous polymeric scaffolding system forin vitrosimulation of pancreatic ductal adenocarcinoma

“…While 2D in vitro systems are easy to use and responsive to radiation and drugs 8,16,17 they cannot accurately simulate important 3D in vivo TME aspects such as structure, porosity, presence of ECM proteins, realistic cell-cell and cell-ECM interactions, environmental gradients (nutrient, oxygen) and vascularisation. 8,[18][19][20][21] Several animal model systems have been developed for pancreatic cancer, 22,23 the best of which involve genetic modication, that have provided considerable insights into the pathogenesis of the disease. However, they are time consuming, expensive to generate and maintain and they are not always reproducible.…”

In Vitro 3D Cultures to Reproduce the Bone Marrow Niche

“…Glucose was found to have a greater impact on cell growth in the 3D model. These observations could be used to facilitate treatment optimization …”

“…These observations could be used to facilitate treatment optimization. (95) Bone-metastatic breast cancer Along with primary BM cancer, metastatic disease comprises a significant portion of malignancy involving the marrow, often with greater morbidity and mortality. Of the cancers that commonly metastasize to bone, here we will focus on breast cancer and PCa, which comprise 80% of bone metastatic disease.…”