Characterization of an Alginate Encapsulated LS180 Spheroid Model for Anti-colorectal Cancer Compound Screening

Abstract

Colorectal cancer is one of the leading causes of cancer-related deaths. A main problem for its treatment is resistance to chemotherapy, requiring the development of new drugs. The success rate of new candidate cancer drugs in clinical trials remains dismal. Three-dimensional (3D) cell culture models have been proposed to bridge the current gap between in vitro chemotherapeutic studies and the human in vivo, due to shortcomings in the physiological relevance of the commonly used two-dimensional cell culture models. In this study, LS180 colorectal cancer cells were cultured as 3D sodium alginate encapsulated spheroids in clinostat bioreactors. Growth and viability were evaluated for 20 days to determine the ideal experimental window. The 3- (4,5- dimethylthiazol- 2- yl)-2,5-diphenyltetrazolium bromide assay was then used to establish half maximal inhibitory concentrations for the standard chemotherapeutic drug, paclitaxel. This concentration was used to further evaluate the established 3D model. During model characterization and evaluation soluble protein content, intracellular adenosine triphosphate levels, extracellular adenylate kinase, glucose consumption, and P-glycoprotein (P-gp) gene expression were measured. Use of the model for chemotherapeutic treatment screening was evaluated using two concentrations of paclitaxel, and treatment continued for 96 h. Paclitaxel caused a decrease in cell growth, viability, and glucose consumption in the model. Furthermore, relative expression of P-gp increased compared to the untreated control group. This is a typical resistance-producing change, seen in vivo and known to be a result of paclitaxel treatment. It was concluded that the LS180 sodium alginate encapsulated spheroid model could be used for testing new chemotherapeutic compounds for colorectal cancer.

A 3D View of Colorectal Cancer Models in Predicting Therapeutic Responses and Resistance

“…Alginate beads containing HCT116 cells, a CRC cell line, have been previously used to demonstrate relative drug resistance and the effects of fluorouracil (5-FU) chemotherapy on CRC cells in 3D [ 125 ]. Treating HCT116 in alginate gels with paclitaxel showed a decrease in glucose uptake, cell proliferation and cell viability [ 126 ]. Both of these studies concluded that embedding CRC cells in alginate beads could be used to test chemotherapeutic drugs.…”

A Biomimetic Model for Liver Cancer to Study Tumor-Stroma Interactions in a 3D Environment with Tunable Bio-Physical Properties

“…Based on the previous experience of working with hydrogels an experiment was setup to determine the optimal cell seeding density (data not shown) 37 . Cells were embedded into a combination of collagen and fibrinogen in the following concentration range (7.5 x 10 5 , 8.5 x 10 5 , 9.5 x 10 5 , 1.0 x 10 6 , 1.5 x 10 6 and 2.0 x 10 6 cells/mL), 2.0 x 10 6 cells/mL was found to be the optimal seeding density.…”

Advanced 3D Cell Culture Techniques in Micro-Bioreactors, Part II: Systems and Applications

“…Other applications of the RWV technique comprise tissue engineering approaches for functional cardiac muscle from rat heart cells [64] or from mouse embryonic stem cells [65] as well as bone tissue engineering [66,67], chondrogenesis [68], connective tissue [69], pancreas [70], and Schwann cells [71]. A human vaginal epithelial cell model [72], a cytotrophoblast invasion model [73], and a human intestinal epithelial cells (INT-407) for implantation model [74] and human colorectal cancer cell lines were used (LS180) for compound screening [75]. More general studies such as that of Wrzesinski et al showed that metabolic reprogramming of 3D cultures is strongly influenced by hypoxia [76] and Chang et al studied the molecular mechanisms underlying the enhanced functions of 3D hepatocyte aggregates [62].…”