A novel three‑dimensional tumorsphere culture system for the efficient and low‑cost enrichment of cancer stem cells with natural polymers

Abstract

Cancer stem cells (CSCs) are considered to serve a key role in tumor progression, recurrence and metastasis. Tumorsphere culture is the most important method for enriching CSCs and is widely used in basic research and drug screening. However, the traditional suspension cell culture system has several disadvantages, including low efficiency, high cost and difficult procedure, making it difficult to produce tumorspheres on a large scale. In the present study, two biomaterials, methylcellulose (MC) and gellan gum (GG), were used to construct a novel culture system based on the traditional system. Subsequently, the characteristics of the novel three-dimensional (3D) culture system were evaluated, the design scheme was optimized, and the morphological and biological features of the tumorspheres cultured in this 3D system were compared with the traditional system. The results revealed that the tumorspheres cultured in the novel 3D system presented a higher seeding density and improved morphology, while maintaining stem-like properties. This evidence suggests that a simple, efficient and low-cost culture system that produces tumorspheres on a large scale was successfully constructed, which can be widely used in various aspects of stem cell research.

Pleurotus highking mushrooms potentiate antiproliferative and antimigratory activity against triple-negative breast cancer cells by suppressing Akt signaling

“…In vitro tumor-sphere formation is a new and inexpensive method considered a potential alternative for in vivo experiments and is now frequently used for the screening of anticancer drugs. 17 In the present study, the number and size of tumor spheres were sharply reduced by PEF-III treatment in a concentration-dependent manner. The mRNA profile of PEF-III-treated tumor-sphere cells showed that PEF-III treatment clearly suppressed the proliferation gene and expression of metastasis-modulating genes.…”

PD-L1 Expression in Human Breast Cancer Stem Cells Is Epigenetically Regulated through Posttranslational Histone Modifications

“…It has been reported that tumorsphere culture system provides a useful method for maintaining a CSC microenvironment [18]. In this study, we enriched the cells having stem cell property in MCF-7 and BT-549 cell lines using tumorsphere formation assay.…”

A Marine Collagen-Based Biomimetic Hydrogel Recapitulates Cancer Stem Cell Niche and Enhances Progression and Chemoresistance in Human Ovarian Cancer

“…Moreover, numerous challenges remain with the traditional suspension culture system. These include low yield and difficulty in maintaining spheroids and changing the media procedure [ 90 , 91 , 103 ].…”

“…Second, different scaffold-based 3D cell culture techniques that use various types of scaffolds have recently been reported to develop advanced methods for CSC enrichment. These include chitosan-alginate scaffolds [ 104 ], polyelectrolyte multilayer films [ 105 ], hyaluronic acid-based multilayer films [ 106 ], porous chitosan-alginate scaffolds [ 107 ], poly (ε-caprolactone) scaffolds [ 108 ], Matrigel ® [ 109 ], agarose multi-well dishes [ 110 ], methylcellulose and gellan gum [ 103 ], and hyaluronic acid hydrogels [ 111 ]. Despite recent advancements in CSC-enrichment techniques, improving the methods for enrichment of the desired CSCs remains an important challenge.…”