In vitro characterization of spheres derived from colorectal cancer cell lines

Olejniczak, Agata; Szaryńska, Magdalena; Kmieć, Zbigniew

doi:10.3892/ijo.2017.4206

Cited by 29 publications

(48 citation statements)

References 71 publications

(91 reference statements)

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“…4B). The untreated HT29 cell-derived spheres presented a lower percentage of CD133 + cells than the HCT116 cell-derived spheres, as we have reported previously (10). Phenotypic analysis revealed a statistically significant decrease in the fraction of HT29-CD133 + only in the 5-FU + ASA-treated cells (Fig.…”

Section: Resultssupporting

confidence: 82%

“…The HT29-derived colonospheres presented a lower rate of proliferation in comparison to the HCT116-derived colonospheres ( Fig. 9D), as we have reported previously (10). Moreover, the HT29 cells cultured with the agents displayed varying patterns of cellular division than the HCT116-derived colonospheres, since the HT29-derived colonospheres exhibited a lower percentage of dividing cells following treatment (Fig.…”

Section: Effects Of the Tested Agents On The Proliferation And Cell Csupporting

confidence: 78%

“…Phenotypic analysis of colonospheres following drug treatment. The phenotypic analysis of the drug-treated spheres aimed to determine the fractions of putative CSC-like cells that express characteristic markers, such as CD133, CD29 and CD44 (10,20). We observed a significant decrease in the percentage of CD133 + cells, a marker which was for the first time used to define CSC-like cells from CRC (2,3), in HCT116 cell-derived spheres following incubation with the majority of the applied drugs.…”

Section: Resultsmentioning

confidence: 85%

“…with ASA on the spherical culture system of HCT116 and HT29 colorectal cancer cell lines cell cultures are unable to mimic the natural elements of the tumor spatial organization. Hence, in recent years, spherical models of cancer cell cultures have gained popularity, as they more adequately represent tumor growth and in vivo development (2,3,8,10,11). Therefore, a crucial process for the effective testing of anticancer drugs is the development of novel in vitro cell culture systems that can better reproduce such traits of in vivo tumors as varied expression of genes, access to nutrients, oxygen, growth factors or waste metabolites delivery systems (8).…”

Section: Effects Of 5-fu and Anti-egfr Antibody In Combinationmentioning

confidence: 99%

“…However, CSC-like cells represent a diversified population which can reside in multiple, alternative phenotypic states within a tumor, accounting for intratumoral cell heterogeneity (16,17). Although cell sorting can enrich the cancer cell population with cells possessing tumor-initiating capacity, the expression of CSC-markers is variable, not only among patient samples, but also between established cancer cell lines (2,3,10,18,19). Independently of some discrepancies in the estimations of the number of CSC-like cells within spherical cell culture systems, it can be claimed that spherical models present a useful tool for testing the effectiveness of anticancer agents.…”

Section: Effects Of 5-fu and Anti-egfr Antibody In Combinationmentioning

confidence: 99%

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Effects of 5-FU and anti-EGFR antibody in combination with ASA on the spherical culture system of HCT116 and HT29 colorectal cancer cell lines

et al. 2019

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The aim of this study was to examine the effects of 5-fluorouracil (5-FU), anti-epidermal growth factor receptor (EGFR) antibody and aspirin (ASA) on the characteristics of two CRC cell lines, HCT116 and HT29, maintained in a spherical culture system. We observed that the morphology of both the HCT116 and HT29 cell-derived spheres was significantly impaired and the size of the colonospheres was markedly reduced following treatment with the aforementioned three drugs. In contrast to adherent cultures, the spherical cultures were more resistant to the tested drugs, as was reflected by their capacity to recreate the colonospheres when sustained in serum-free medium. Flow cytometric analysis of the drug-treated HCT116 cell-derived spheres revealed changes in the fraction of cells expressing markers of cancer stem cells (CSCs), whereas the CSC phenotype of HT29 cell-derived colonospheres was affected to a lesser extent. All reagents enhanced the percentage of non-viable cells in the colonospheres despite the diminished fraction of active caspase-3-positive cells following treatment of the HT29 cell-derived spheres with anti-EGFR antibody. Increased autophagy, assessed by acridine orange staining, was noted following the incubation of the HT29-colonospheres with ASA and 5-FU in comparison to the control. Notably, the percentage of cyclooxygenase (COX)-2-positive cells was not affected by ASA, although its activity was markedly elevated in the colonospheres incubated with anti-EGFR antibody. On the whole, the findings of this study indicate that all the tested drugs were involved in different cellular processes, which suggests that they should be considered for the combined therapeutic treatment of CRC, particularly for targeting the population of CSC-like cells. Thus, cancer cell-derived spheres may be used as a preferable model for in vitro anticancer drug testing.

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Section: Resultssupporting

confidence: 82%

Section: Effects Of the Tested Agents On The Proliferation And Cell Csupporting

confidence: 78%

Section: Resultsmentioning

confidence: 85%

Section: Effects Of 5-fu and Anti-egfr Antibody In Combinationmentioning

confidence: 99%

Section: Effects Of 5-fu and Anti-egfr Antibody In Combinationmentioning

confidence: 99%

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Effects of 5-FU and anti-EGFR antibody in combination with ASA on the spherical culture system of HCT116 and HT29 colorectal cancer cell lines

et al. 2019

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Establishment of a tissue‐engineered colon cancer model for comparative analysis of cancer cell lines

Hassani,

Anbiah,

Moore

et al. 2023

J Biomedical Materials Res

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To overcome the limitations of in vitro two‐dimensional (2D) cancer models in mimicking the complexities of the native tumor milieu, three‐dimensional (3D) engineered cancer models using biomimetic materials have been introduced to more closely recapitulate the key attributes of the tumor microenvironment. Specifically, for colorectal cancer (CRC), a few studies have developed 3D engineered tumor models to investigate cell–cell interactions or efficacy of anti‐cancer drugs. However, recapitulation of CRC cell line phenotypic differences within a 3D engineered matrix has not been systematically investigated. Here, we developed an in vitro 3D engineered CRC (3D‐eCRC) tissue model using the natural‐synthetic hybrid biomaterial PEG‐fibrinogen and three CRC cell lines, HCT 116, HT‐29, and SW480. To better recapitulate native tumor conditions, our 3D‐eCRC model supported higher cell density encapsulation (20 × 106 cells/mL) and enabled longer term maintenance (29 days) as compared to previously reported in vitro CRC models. The 3D‐eCRCs formed using each cell line demonstrated line‐dependent differences in cellular and tissue properties, including cellular growth and morphology, cell subpopulations, cell size, cell granularity, migration patterns, tissue growth, gene expression, and tissue stiffness. Importantly, these differences were found to be most prominent from Day 22 to Day 29, thereby indicating the importance of long‐term culture of engineered CRC tissues for recapitulation and investigation of mechanistic differences and drug response. Our 3D‐eCRC tissue model showed high potential for supporting future in vitro comparative studies of disease progression, metastatic mechanisms, and anti‐cancer drug candidate response in a CRC cell line‐dependent manner.

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Enrichment of Melanoma Stem-Like Cells via Sphere Assays

Mukherjee

Lambert

Norris

et al. 2021

Methods in Molecular Biology

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Sphere assays are widely used in vitro techniques to enrich and evaluate the stem-like cell behavior of both normal and cancer cells. Utilizing three-dimensional in vitro sphere culture conditions provide a better representation of tumor growth in vivo than the more common monolayer cultures. We describe how to perform primary and secondary sphere assays, used for the enrichment and self-renewability studies of melanoma/melanocyte stem-like cells. Spheres are generated by growing melanoma cells at low density in nonadherent conditions with stem cell media. We provide protocols for preparing inexpensive and versatile polyHEMA-coated plates, setting up primary and secondary sphere assays in almost any tissue culture format and quantification methods using standard inverted microscopy. Our protocol is easily adaptable to laboratories with basic cell culture capabilities, without the need for expensive fluidic instruments.

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In vitro characterization of spheres derived from colorectal cancer cell lines

Cited by 29 publications

References 71 publications

Effects of 5-FU and anti-EGFR antibody in combination with ASA on the spherical culture system of HCT116 and HT29 colorectal cancer cell lines

Effects of 5-FU and anti-EGFR antibody in combination with ASA on the spherical culture system of HCT116 and HT29 colorectal cancer cell lines

Establishment of a tissue‐engineered colon cancer model for comparative analysis of cancer cell lines

Enrichment of Melanoma Stem-Like Cells via Sphere Assays

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