Editorial

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Open AccessEditorial

Three-Dimensional (3D) Culture Models in Cancer Investigation, Drug Testing and Immune Response Evaluation

by Roberto Benelli, Maria Raffaella Zocchi and Alessandro Poggi

Int. J. Mol. Sci. 2021, 22(1), 150; https://doi.org/10.3390/ijms22010150 - 25 Dec 2020

Cited by 2 | Viewed by 1805

Preclinical models for the definition of anti-cancer drug safety and efficacy are constantly evolving [...] Full article

(This article belongs to the Special Issue Application of 3D Culture Systems in Cancer Development and Anti-tumor Drug Selection: Advantages and Drawbacks)

Research

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17 pages, 2348 KiB

Open AccessArticle

Digital Image Analysis Applied to Tumor Cell Proliferation, Aggressiveness, and Migration-Related Protein Synthesis in Neuroblastoma 3D Models

by Ezequiel Monferrer, Sabina Sanegre, Susana Martín-Vañó, Andrea García-Lizarribar, Rebeca Burgos-Panadero, Amparo López-Carrasco, Samuel Navarro, Josep Samitier and Rosa Noguera

Int. J. Mol. Sci. 2020, 21(22), 8676; https://doi.org/10.3390/ijms21228676 - 17 Nov 2020

Cited by 6 | Viewed by 3669

Abstract

Patient-derived cancer 3D models are a promising tool that will revolutionize personalized cancer therapy but that require previous knowledge of optimal cell growth conditions and the most advantageous parameters to evaluate biomimetic relevance and monitor therapy efficacy. This study aims to establish general [...] Read more.

Patient-derived cancer 3D models are a promising tool that will revolutionize personalized cancer therapy but that require previous knowledge of optimal cell growth conditions and the most advantageous parameters to evaluate biomimetic relevance and monitor therapy efficacy. This study aims to establish general guidelines on 3D model characterization phenomena, focusing on neuroblastoma. We generated gelatin-based scaffolds with different stiffness and performed SK-N-BE(2) and SH-SY5Y aggressive neuroblastoma cell cultures, also performing co-cultures with mouse stromal Schwann cell line (SW10). Model characterization by digital image analysis at different time points revealed that cell proliferation, vitronectin production, and migration-related gene expression depend on growing conditions and are specific to the tumor cell line. Morphometric data show that 3D in vitro models can help generate optimal patient-derived cancer models, by creating, identifying, and choosing patterns of clinically relevant artificial microenvironments to predict patient tumor cell behavior and therapeutic responses. Full article

(This article belongs to the Special Issue Application of 3D Culture Systems in Cancer Development and Anti-tumor Drug Selection: Advantages and Drawbacks)

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18 pages, 5531 KiB

Open AccessArticle

Rapid Clearing for High Resolution 3D Imaging of Ex Vivo Pancreatic Cancer Spheroids

by Eliana Steinberg, Natalie Orehov, Katerina Tischenko, Ouri Schwob, Gideon Zamir, Ayala Hubert, Zakhariya Manevitch and Ofra Benny

Int. J. Mol. Sci. 2020, 21(20), 7703; https://doi.org/10.3390/ijms21207703 - 18 Oct 2020

Cited by 10 | Viewed by 3971

Abstract

The currently accepted imaging methods have been a central hurdle to imaging the finer details of tumor behavior in three-dimensional (3D) ex vivo multicellular culture models. In our search for an improved way of imaging tumor behavior in its physiological-like niche, we developed [...] Read more.

The currently accepted imaging methods have been a central hurdle to imaging the finer details of tumor behavior in three-dimensional (3D) ex vivo multicellular culture models. In our search for an improved way of imaging tumor behavior in its physiological-like niche, we developed a simple, efficient, and straightforward procedure using standard reagents and imaging equipment that significantly enhanced 3D imaging up to a ~200-micron depth. We tested its efficacy on pancreatic spheroids, prototypes of high-density tissues that are difficult to image. We found we could both save time with this method and extract information about pancreatic tumor spheroids that previously was difficult to obtain. We were able to discern clear differences in the organization of pancreatic tumor spheroids generated from different origins, suggesting cell-specific, inherent, bottom-up organization with a correlation to the level of malignancy. We also examined the dynamic changes in the spheroids at predetermined time points, providing important information related to tissue morphogenesis and its metabolic state. Lastly, this process enabled us to assess a drug vehicle’s potential to penetrate dense tumor tissue by improving our view of the inert particles’ diffusion in the 3D spheroid. This clearing method, a simple procedure, can open the door to more accurate imaging and reveal more about cancer behavior. Full article

(This article belongs to the Special Issue Application of 3D Culture Systems in Cancer Development and Anti-tumor Drug Selection: Advantages and Drawbacks)

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18 pages, 4754 KiB

Open AccessArticle

Astrocytes Decreased the Sensitivity of Glioblastoma Cells to Temozolomide and Bay 11-7082

by Sadaf E. Pustchi, Naze G. Avci, Yasemin M. Akay and Metin Akay

Int. J. Mol. Sci. 2020, 21(19), 7154; https://doi.org/10.3390/ijms21197154 - 28 Sep 2020

Cited by 11 | Viewed by 3654

Abstract

Glioblastoma multiforme (GBM) is the most common malignant type of astrocytic tumors. GBM patients have a poor prognosis with a median survival of approximately 15 months despite the “Stupp” Regimen and high tumor recurrence due to the tumor resistance to chemotherapy. In this [...] Read more.

Glioblastoma multiforme (GBM) is the most common malignant type of astrocytic tumors. GBM patients have a poor prognosis with a median survival of approximately 15 months despite the “Stupp” Regimen and high tumor recurrence due to the tumor resistance to chemotherapy. In this study, we co-cultured GBM cells with human astrocytes in three-dimensional (3D) poly(ethylene glycol) dimethyl acrylate (PEGDA) microwells to mimic the tumor microenvironment. We treated 3D co- and mono-cultured cells with Temozolomide (TMZ) and the nuclear factor-κB (NF-κB) inhibitor Bay 11-7082 and investigated the combined effect of the drugs. We assessed the expressions of glial fibrillary acidic protein (GFAP) and vimentin that play a role in the tumor malignancy and activation of the astrocytes as well as Notch-1 and survivin that play a role in GBM malignancy after the drug treatment to understand how astrocytes induced GBM drug response. Our results showed that in the co-culture, astrocytes increased GBM survival and resistance after combined drug treatment compared to mono-cultures. These data restated the importance of 3D cell culture to mimic the tumor microenvironment for drug screening. Full article

(This article belongs to the Special Issue Application of 3D Culture Systems in Cancer Development and Anti-tumor Drug Selection: Advantages and Drawbacks)

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16 pages, 3609 KiB

Open AccessArticle

Assessing Advantages and Drawbacks of Rapidly Generated Ultra-Large 3D Breast Cancer Spheroids: Studies with Chemotherapeutics and Nanoparticles

by Austin R. Holub, Anderson Huo, Kavil Patel, Vishal Thakore, Pranav Chhibber and Folarin Erogbogbo

Int. J. Mol. Sci. 2020, 21(12), 4413; https://doi.org/10.3390/ijms21124413 - 21 Jun 2020

Cited by 9 | Viewed by 4870

Abstract

Traditionally, two-dimensional (2D) monolayer cell culture models have been used to study in vitro conditions for their ease of use, simplicity and low cost. However, recently, three-dimensional (3D) cell culture models have been heavily investigated as they provide better physiological relevance for studying [...] Read more.

Traditionally, two-dimensional (2D) monolayer cell culture models have been used to study in vitro conditions for their ease of use, simplicity and low cost. However, recently, three-dimensional (3D) cell culture models have been heavily investigated as they provide better physiological relevance for studying various disease behaviors, cellular activity and pharmaceutical interactions. Typically, small-sized tumor spheroid models (100–500 μm) are used to study various biological and physicochemical activities. Larger, millimetric spheroid models are becoming more desirable for simulating native tumor microenvironments (TMEs). Here, we assess the use of ultra-large spheroid models (~2000 μm) generated from scaffolds made from a nozzle-free, ultra-high resolution printer; these models are explored for assessing chemotherapeutic responses with molecular doxorubicin (DOX) and two analogues of Doxil^Ⓡ (Dox-NP^Ⓡ, Doxoves^TM) on MDA-MB-231 and MCF-7 breast cancer cell lines. To provide a comparative baseline, small spheroid models (~500 μm) were developed using a self-aggregation method of MCF-7 breast cancer cell lines, and underwent similar drug treatments. Analysis of both large and small MCF-7 spheroids revealed that Dox-NP tends to have the highest level of inhibition, followed by molecular doxorubicin and then Doxoves. The experimental advantages and drawbacks of using these types of ultra-large spheroids for cancer research are discussed. Full article

(This article belongs to the Special Issue Application of 3D Culture Systems in Cancer Development and Anti-tumor Drug Selection: Advantages and Drawbacks)

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23 pages, 4295 KiB

Open AccessArticle

Evaluation of Melanoma (SK-MEL-2) Cell Growth between Three-Dimensional (3D) and Two-Dimensional (2D) Cell Cultures with Fourier Transform Infrared (FTIR) Microspectroscopy

by Tarapong Srisongkram, Natthida Weerapreeyakul and Kanjana Thumanu

Int. J. Mol. Sci. 2020, 21(11), 4141; https://doi.org/10.3390/ijms21114141 - 10 Jun 2020

Cited by 14 | Viewed by 4334

Abstract

Fourier transform infrared (FTIR) microspectroscopy was used to evaluate the growth of human melanoma cells (SK-MEL-2) in two-dimensional (2D) versus three-dimensional (3D) spheroid culture systems. FTIR microspectroscopy, coupled with multivariate analysis, could be used to monitor the variability of spheroid morphologies prepared from [...] Read more.

Fourier transform infrared (FTIR) microspectroscopy was used to evaluate the growth of human melanoma cells (SK-MEL-2) in two-dimensional (2D) versus three-dimensional (3D) spheroid culture systems. FTIR microspectroscopy, coupled with multivariate analysis, could be used to monitor the variability of spheroid morphologies prepared from different cell densities. The characteristic shift in absorbance bands of the 2D cells were different from the spectra of cells from 3D spheroids. FTIR microspectroscopy can also be used to monitor cell death similar to fluorescence cell staining in 3D spheroids. A change in the secondary structure of protein was observed in cells from the 3D spheroid versus the 2D culture system. FTIR microspectroscopy can detect specific alterations in the biological components inside the spheroid, which cannot be detected using fluorescence cell death staining. In the cells from 3D spheroids, the respective lipid, DNA, and RNA region content represent specific markers directly proportional to the spheroid size and central area of necrotic cell death, which can be confirmed using unsupervised PCA and hierarchical cluster analysis. FTIR microspectroscopy could be used as an alternative tool for spheroid cell culture discrimination, and validation of the usual biochemical technique. Full article

(This article belongs to the Special Issue Application of 3D Culture Systems in Cancer Development and Anti-tumor Drug Selection: Advantages and Drawbacks)

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18 pages, 3057 KiB

Open AccessArticle

Synergy between Photodynamic Therapy and Dactinomycin Chemotherapy in 2D and 3D Ovarian Cancer Cell Cultures

by Layla Mohammad Hadi, Elnaz Yaghini, Alexander J. MacRobert and Marilena Loizidou

Int. J. Mol. Sci. 2020, 21(9), 3203; https://doi.org/10.3390/ijms21093203 - 30 Apr 2020

Cited by 13 | Viewed by 3490

Abstract

In this study we explored the efficacy of combining low dose photodynamic therapy using a porphyrin photosensitiser and dactinomycin, a commonly used chemotherapeutic agent. The studies were carried out on compressed collagen 3D constructs of two human ovarian cancer cell lines (SKOV3 and [...] Read more.

In this study we explored the efficacy of combining low dose photodynamic therapy using a porphyrin photosensitiser and dactinomycin, a commonly used chemotherapeutic agent. The studies were carried out on compressed collagen 3D constructs of two human ovarian cancer cell lines (SKOV3 and HEY) versus their monolayer counterparts. An amphiphilc photosensitiser was employed, disulfonated tetraphenylporphine, which is not a substrate for ABC efflux transporters that can mediate drug resistance. The combination treatment was shown to be effective in both monolayer and 3D constructs of both cell lines, causing a significant and synergistic reduction in cell viability. Compared to dactinomycin alone or PDT alone, higher cell kill was found using 2D monolayer culture vs. 3D culture for the same doses. In 3D culture, the combination therapy resulted in 10 and 22 times higher cell kill in SKOV3 and HEY cells at the highest light dose compared to dactinomycin monotherapy, and 2.2 and 5.5 times higher cell kill than PDT alone. The combination of low dose PDT and dactinomycin appears to be a promising way to repurpose dactinomycin and widen its therapeutic applications. Full article

(This article belongs to the Special Issue Application of 3D Culture Systems in Cancer Development and Anti-tumor Drug Selection: Advantages and Drawbacks)

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14 pages, 5840 KiB

Open AccessArticle

3D Bioprinted Vascularized Tumour for Drug Testing

by Seokgyu Han, Sein Kim, Zhenzhong Chen, Hwa Kyoung Shin, Seo-Yeon Lee, Hyo Eun Moon, Sun Ha Paek and Sungsu Park

Int. J. Mol. Sci. 2020, 21(8), 2993; https://doi.org/10.3390/ijms21082993 - 23 Apr 2020

Cited by 55 | Viewed by 7251

Abstract

An in vitro screening system for anti-cancer drugs cannot exactly reflect the efficacy of drugs in vivo, without mimicking the tumour microenvironment (TME), which comprises cancer cells interacting with blood vessels and fibroblasts. Additionally, the tumour size should be controlled to obtain reliable [...] Read more.

An in vitro screening system for anti-cancer drugs cannot exactly reflect the efficacy of drugs in vivo, without mimicking the tumour microenvironment (TME), which comprises cancer cells interacting with blood vessels and fibroblasts. Additionally, the tumour size should be controlled to obtain reliable and quantitative drug responses. Herein, we report a bioprinting method for recapitulating the TME with a controllable spheroid size. The TME was constructed by printing a blood vessel layer consisting of fibroblasts and endothelial cells in gelatine, alginate, and fibrinogen, followed by seeding multicellular tumour spheroids (MCTSs) of glioblastoma cells (U87 MG) onto the blood vessel layer. Under MCTSs, sprouts of blood vessels were generated and surrounding MCTSs thereby increasing the spheroid size. The combined treatment involving the anti-cancer drug temozolomide (TMZ) and the angiogenic inhibitor sunitinib was more effective than TMZ alone for MCTSs surrounded by blood vessels, which indicates the feasibility of the TME for in vitro testing of drug efficacy. These results suggest that the bioprinted vascularized tumour is highly useful for understanding tumour biology, as well as for in vitro drug testing. Full article

(This article belongs to the Special Issue Application of 3D Culture Systems in Cancer Development and Anti-tumor Drug Selection: Advantages and Drawbacks)

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Review

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19 pages, 620 KiB

Open AccessReview

Three-Dimensional Cell Cultures as an In Vitro Tool for Prostate Cancer Modeling and Drug Discovery

by Fabrizio Fontana, Michela Raimondi, Monica Marzagalli, Michele Sommariva, Nicoletta Gagliano and Patrizia Limonta

Int. J. Mol. Sci. 2020, 21(18), 6806; https://doi.org/10.3390/ijms21186806 - 16 Sep 2020

Cited by 33 | Viewed by 8582

Abstract

In the last decade, three-dimensional (3D) cell culture technology has gained a lot of interest due to its ability to better recapitulate the in vivo organization and microenvironment of in vitro cultured cancer cells. In particular, 3D tumor models have demonstrated several different [...] Read more.

In the last decade, three-dimensional (3D) cell culture technology has gained a lot of interest due to its ability to better recapitulate the in vivo organization and microenvironment of in vitro cultured cancer cells. In particular, 3D tumor models have demonstrated several different characteristics compared with traditional two-dimensional (2D) cultures and have provided an interesting link between the latter and animal experiments. Indeed, 3D cell cultures represent a useful platform for the identification of the biological features of cancer cells as well as for the screening of novel antitumor agents. The present review is aimed at summarizing the most common 3D cell culture methods and applications, with a focus on prostate cancer modeling and drug discovery. Full article

(This article belongs to the Special Issue Application of 3D Culture Systems in Cancer Development and Anti-tumor Drug Selection: Advantages and Drawbacks)

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