Comparative analysis of tumor spheroid generation techniques for differential<i>in vitro</i>drug toxicity

Raghavan, Shreya; Mehta, Pooja; Horst, Eric N.; Ward, Maria R.; Rowley, Katelyn R.; Mehta, Geeta

doi:10.18632/oncotarget.7659

Cited by 136 publications

(159 citation statements)

References 32 publications

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“…Cisplatin Breast cancer EMT-6 [24] Endometrial cancer Ishikawa [25] RL95-2 [25] KLE [25] Hypopharyngeal cancer FaDu [96] Ovarian cancer OVCAR8 [97] Oral cancer SCC25 [96] CAL27 [96] Chlorimipramine Glioma C6 [98] Cyclophosphamide Breast cancer EMT-6 [24] Diclofenac Liver tissue Hepatocytes obtained from patient [10] Hepatoma C3A [68] Docetaxel Cervical cancer HeLa [99] Colon cancer HT29 [99] Doxorubicin Cervical cancer HeLa [70] Endometrial cancer Ishikawa [25] RL95-2 [25] KLE [25] Hepatoma C3A [68] Huh7 [100] Lung cancer A549 [101] E-combretastatin A4 Melanoma C8161 [102] Breast cancer MDA 231 [102] (Continued) [26] PC3 [26] Fialuridine Liver tissue Hepatocytes obtained from patient [10] Fialuridine Hepatoma C3A [68] Gant61…”

Section: Cb13mentioning

confidence: 99%

Spheroids Formation on Non‐Adhesive Surfaces by Liquid Overlay Technique: Considerations and Practical Approaches

Costa

Melo‐Diogo

Moreira

et al. 2017

Biotechnology Journal

130

125

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Scalable and reproducible production of 3D cellular spheroids is highly demanded, by pharmaceutical companies, for drug screening purposes during the pre-clinical evaluation phase. These 3D cellular constructs, unlike the monolayer culture of cells, can mimic different features of human tissues, including cellular organization, cell-cell and cell-extracellular matrix (ECM) interactions. Up to now, different techniques (scaffold-based and -free) have been used for spheroids formation, being the Liquid Overlay Technique (LOT) one of the most explored methodologies, due to its low cost and easy handling. Additionally, during the last few decades, this technique has been widely investigated in order to enhance its potential for being applied in high-throughput analysis. Herein, an overview of the LOT advances, practical approaches, and troubleshooting is provided for those researchers that intend to produce spheroids using LOT, for drug screening purposes. Moreover, the advantages of the LOT over the other scaffold-free techniques used for the spheroids formation are also addressed. Highlights • 2D cell culture drawbacks are summarized;• spheroids mimic the features of human tissues;• scaffold-based and scaffold-free technologies for spheroids production are discussed; • advantages of LOT over other scaffold-free techniques are highlighted; • LOT advances, practical approaches and troubleshooting are underlined.

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Section: Cb13mentioning

confidence: 99%

Spheroids Formation on Non‐Adhesive Surfaces by Liquid Overlay Technique: Considerations and Practical Approaches

Costa

Melo‐Diogo

Moreira

et al. 2017

Biotechnology Journal

130

125

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“…They closely resemble small avascular tumors with complex cell–cell and cell–matrix interactions 9,10. In addition, MCTS is a powerful 3D in vitro model to study tumor and tumor cell proliferation, morphology, cell invasion, drug binding, and chemo-resistance 11–14. Compared with two dimensional (2D) monolayer cell platforms, MCTS expresses increased resistance to chemotherapy, perhaps due to their ability to mimic multicellular arrangements and ECM deposition of avascular tumors 15.…”

Section: Introductionmentioning

confidence: 99%

Sialylation facilitates self-assembly of 3D multicellular prostaspheres by using cyclo-RGDfK(TPP) peptide

Haq¹,

Samuel²,

Haxho³

et al. 2017

OTT

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BackgroundProstaspheres-based three dimensional (3D) culture models have provided insight into prostate cancer (PCa) biology, highlighting the importance of cell–cell interactions and the extracellular matrix (EMC) in the tumor microenvironment. Although these 3D classical spheroid platforms provide a significant advance over 2D models mimicking in vivo tumors, the limitations involve no control of assembly and structure with only limited spatial or glandular organization. Here, matrix-free prostaspheres from human metastatic prostate carcinoma PC3 and DU145 cell lines and their respective gemcitabine resistant (GemR) variants were generated by using cyclic Arg-Gly-Asp-D-Phe-Lys peptide modified with 4-carboxybutyl-triphenylphosphonium bromide (cyclo-RGDfK(TPP)).Materials and methodsMicroscopic imaging, immunocytochemistry (ICC), flow cytometry, sialidase, and WST-1 cell viability assays were used to evaluate the formation of multicellular tumor spheroid (MCTS), cell survival, morphologic changes, and expression levels of α2,6 and α2,3 sialic acid (SA) and E- and N-cadherin in DU145, PC3, and their GemR variants.ResultsBy using the cyclo-RGDfK(TPP) peptide platform in a dose- and time-dependent manner, both DU145 and DU145GemR cells formed small MCTS. In contrast, PC3 and PC3GemR cells formed irregular multicellular aggregates at all concentrations of cyclo-RGDfK(TPP) peptide, even after 6 days of incubation. ICC and flow cytometry results revealed that DU145 cells expressed higher amounts of E-cadherin but lower N-cadherin compared with PC3 cells. By using Maackia amurensis (α2,3-SA-specific MAL-II) and Sambucus nigra (α2,6-SA specific SNA) lectin-based cytochemistry staining and flow cytometry, it was found that DU145 and DU145GemR cells expressed 5 times more α2,6-SA than α2,3-SA on the cell surface. PC3 cells expressed 4 times more α2,3-SA than α2,6-SA, and the PC3GemR cells showed 1.4 times higher α2,6-SA than α2,3-SA. MCTS volume was dose-dependently reduced following pretreatment with α2,6-SA-specific neuraminidase (Vibrio cholerae). Oseltamivir phosphate enhanced cell aggregation and compaction of 3D MCTS formed with PC3 cells.ConclusionThe relative levels of specific sialoglycan structures on the cell surface correlate with the ability of PCa cells to form avascular multicellular prostaspheres.

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“…With this combinatory approach, it will be possible to gain a more complete picture of the cancer microenvironment and ascertain potential avenues of treatment. Additionally, non-cell factors such as chemotaxis, 130 3D culture 94,[131][132][133][134][135][136] , and hypoxia 137 should be considered for future investigations, in conjunction with mechanical cues to create a microenvironment that can more fully recapitulate in vivo conditions. The study of ovarian cancer mechanotransduction promises to improve patient treatment through future investigations that utilize designs pertinent to the specific microenvironment.…”

Section: Discussionmentioning

confidence: 99%

Review: Mechanotransduction in ovarian cancer: Shearing into the unknown

2018

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Perspective: The role of mechanobiology in the etiology of brain metastasis APL Bioengineering 2, 031801 (2018) Ovarian cancer remains a deadly diagnosis with an 85% recurrence rate and a 5-year survival rate of only 46%. The poor outlook of this disease has improved little over the past 50 years owing to the lack of early detection, chemoresistance and the complex tumor microenvironment. Within the peritoneal cavity, the presence of ascites stimulates ovarian tumors with shear stresses. The stiff environment found within the tumor extracellular matrix and the peritoneal membrane are also implicated in the metastatic potential and epithelial to mesenchymal transition (EMT) of ovarian cancer. Though these mechanical cues remain highly relevant to the understanding and treatment of ovarian cancers, our current knowledge of their biological processes and their clinical relevance is deeply lacking. Seminal studies on ovarian cancer mechanotransduction have demonstrated close ties between mechanotransduction and ovarian cancer chemoresistance, EMT, enhanced cancer stem cell populations, and metastasis. This review summarizes our current understanding of ovarian cancer mechanotransduction and the gaps in knowledge that exist. Future investigations on ovarian cancer mechanotransduction will greatly improve clinical outcomes via systematic studies that determine shear stress magnitude and its influence on ovarian cancer progression, metastasis, and treatment.

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Comparative analysis of tumor spheroid generation techniques for differentialin vitrodrug toxicity

Cited by 136 publications

References 32 publications

Spheroids Formation on Non‐Adhesive Surfaces by Liquid Overlay Technique: Considerations and Practical Approaches

Spheroids Formation on Non‐Adhesive Surfaces by Liquid Overlay Technique: Considerations and Practical Approaches

Sialylation facilitates self-assembly of 3D multicellular prostaspheres by using cyclo-RGDfK(TPP) peptide

Review: Mechanotransduction in ovarian cancer: Shearing into the unknown

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