In-depth phenotypic characterization of multicellular tumor spheroids: Effects of 5-Fluorouracil

Abstract

MultiCellular Tumor Spheroids (MCTS), which mimic the 3-Dimensional (3D) organization of a tumor, are considered as better models than conventional cultures in 2-Dimensions (2D) to study cancer cell biology and to evaluate the response to chemotherapeutic drugs. A real time and quantitative follow-up of MCTS with simple and robust readouts to evaluate drug efficacy is still missing. Here, we evaluate the chemotherapeutic drug 5-Fluorouracil (5-FU) response on the growth and integrity of MCTS two days after treatment of MCTS and for three colorectal carcinoma cell lines with different cohesive properties (HT29, HCT116 and SW480). We found different sensitivity to 5-FU for the three CRC cell lines, ranging from high (SW480), intermediate (HCT116) and low (HT29) and the same hierarchy of CRC cell lines sensitivity is conserved in 2D. We also evidence that 5-FU has a strong impact on spheroid cohesion, with the apparition of a number of single detaching cells from the spheroid in a 5-FU dose- and cell line-dependent manner. We propose an innovative methodology for the chemosensitivity evaluation in 3D MCTS that recapitulates and regionalizes the 5-FU-induced changes within MCTS over time. These robust phenotypic read-outs could be easily scalable for high-throughput drug screening that may include different types of cancer cells to take into account tumor heterogeneity and resistance to treatment.

Effective Targeting of Colon Cancer Cells with Piperine Natural Anticancer Prodrug Using Functionalized Clusters of Hydroxyapatite Nanoparticles

“…A novel anticancer targeted drug delivery nanoformulation consisting of aggregates of HAPs With spheroids, HAP-Pip7.2, HAP-P-Pip7.2, HAP-Pip9.3, HAP-P-Pip9.3, HAP-Pip9.3-GA, HAP-P-Pip9.3-GA, HAP-Pip9.3-GA-FA, and HAP-P-Pip9.3-GA-FA had visible changes compared to control and free Pip. The main changes were smaller diameter, shrinkage, and defragmentation of spheroids [81]. The nano-particulate-system containing FA is efficient for delivering Pip prodrug and accumulates in spheroids compared to free Pip; thus, anticancer effects are enhanced due to internalized of HAP-Pip9.3-GA-FA and HAP-P-Pip9.3-GA-FA in HCT116 cells spheroids compared to other formulations.…”

“…A novel anticancer targeted drug delivery nanoformulation consisting of aggregates of HAPs loaded with Pip was efficient against HCT116 colon cancer cells in vitro. We measured a high loading [81]. The nano-particulate-system containing FA is efficient for delivering Pip prodrug and accumulates in spheroids compared to free Pip; thus, anticancer effects are enhanced due to internalized of HAP-Pip9.3-GA-FA and HAP-P-Pip9.3-GA-FA in HCT116 cells spheroids compared to other formulations.…”

Artemisinin Derivatives Stimulate DR5-Specific TRAIL-Induced Apoptosis by Regulating Wildtype P53

“…To further confirm the efficacy of DHA for colon cancer cells, we used 3D spheroids grown from HCT116 and DLD-1 cells to mimic the actual tumor microenvironment. Cells grown in 3D are considered as non-vascularized tumor models that better reflect the 3D cell-cell, cell-matrix interactions, and the biochemical environment of in vivo tumor mass compared to 2D cell models [29]. We incubated the 3D spheroids with single or combination treatments for 24 h after the spheroids were established ( Figure S4A,B).…”

“…This means that DHA overcomes DHER resistance and induces more cell death in the 3D HCT116 cell model. interactions, and the biochemical environment of in vivo tumor mass compared to 2D cell models [29]. We incubated the 3D spheroids with single or combination treatments for 24 h after the spheroids were established ( Figure S4A,B).…”

Precise, high-throughput production of multicellular spheroids with a bespoke 3D bioprinter

“…For 3D cell culture to be used for both fundamental studies and drug screening (7) control over additional culture parameters is required. For example, it is known that volumes and shapes of spheroids influences the sensitivity to anti-cancer drugs (9,10) and the ECM mimic that embed spheroids determines the rate of growth and drug responses (11,12). There are a variety of methods for producing multicellular spheroids using either rotation, microfluidics, non-cell adherent surfaces, magnetic levitation or pealing confluent layers of cells off a surface (13)(14)(15), with the use of non-adherent surfaces being the popular approach (2).…”