Development of transferrin targeted NCL-240 micelles and their evaluation using in-vitro 3D cancer cell culture (spheroid) models

The tumor microenvironment plays a critical role in modulating cancer cell migration, metabolism, and malignancy, thus, highlighting the need to develop in vitro culture systems that can recapitulate its abnormal properties. While a variety of stiffness-tunable biomaterials, reviewed here, have been developed to mimic the rigidity of the tumor extracellular matrix, culture systems that can recapitulate the broader extracellular context of the tumor microenvironment (including pH and temperature) remain comparably unexplored, partially due to the difficulty in independently tuning these parameters. Here, we investigate a self-assembled polypeptide network hydrogel as a cell culture platform and demonstrate that the culture parameters, including the substrate stiffness, extracellular pH and temperature, can be independently controlled. We then use this biomaterial as a cell culture substrate to assess the effect of stiffness, pH and temperature on Suit2 cells, a pancreatic cancer cell line, and demonstrate that these microenvironmental factors can regulate two critical transcription factors in cancer: yes-associated protein 1 (YAP) and hypoxia inducible factor (HIF-1A).

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Smart Shockwave Responsive Titania-Based Nanoparticles for Cancer Treatment

Pharmaceutics ◽

10.3390/pharmaceutics13091423 ◽

2021 ◽

Vol 13 (9) ◽

pp. 1423

Author(s):

Veronica Vighetto ◽

Luisa Racca ◽

Marta Canta ◽

Joana C. Matos ◽

Bianca Dumontel ◽

...

Keyword(s):

Cell Culture ◽

Shock Waves ◽

Cancer Cell ◽

Colloidal Stability ◽

Culture Media ◽

High Sensitivity ◽

High Energy ◽

High Dispersion ◽

Specific Chemical

Nanomedicine is an emerging treatment approach for many cancers, characterized by having high sensitivity and selectivity for tumor cells and minimal toxic effects induced by the conventional chemotherapeutics. In these context, smart nanoparticles (NPs) are getting increasingly relevant in the development of new therapies. NPs with specific chemical composition and/or structure and being stimuli-responsive to magnetic, light or ultrasound waves are new promising tools. In the present work, amorphous-titania propyl-amine functionalized (a-TiO2-NH2) NPs, coated with bovine serum albumin (BSA), are stimulated with high energy shock waves to induce cytotoxic effects in cancer cells. First, a new method to coat a-TiO2-NH2 NPs with BSA (a-TiO2-NH2/BSA) was proposed, allowing for a high dispersion and colloidal stability in a cell culture media. The a-TiO2-NH2/BSA NPs showed no cancer cell cytotoxicity. In a second step, the use of shock waves to stimulate a-TiO2-NH2/BSA NPs, was evaluated and optimized. A systematic study was performed in in vitro cell culture aiming to impair the cancer cell viability: NP concentrations, time steps and single versus multiple shock waves treatments were studied. The obtained results highlighted the relevance of NPs design and administration time point with respect to the shock wave treatment and allow to hypothesize mechanical damages to cells.

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Application and research progress of in vitro liver cancer cell culture models

World Chinese Journal of Digestology ◽

10.11569/wcjd.v29.i11.563 ◽

2021 ◽

Vol 29 (11) ◽

pp. 563-570

Author(s):

Jia-Li Xing ◽

Yu-Xin Wang ◽

Shun-Da Du

Keyword(s):

Cell Culture ◽

Liver Cancer ◽

Cancer Cell ◽

Research Progress ◽

Liver Cancer Cell ◽

Culture Models ◽

Cell Culture Models

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Montmorillonite colloid plates with adsorbed cytochrome c: in vitro cytotoxic effect on colon cancer cell culture

Cancer Nanotechnology ◽

10.1186/s12645-021-00095-7 ◽

2021 ◽

Vol 12 (1) ◽

Author(s):

Svetlana H. Hristova ◽

Alexandar M. Zhivkov

Keyword(s):

Cell Culture ◽

Colon Cancer ◽

Cancer Cells ◽

Cancer Cell ◽

Aqueous Suspension ◽

Colon Cancer Cell ◽

Composite Particles ◽

Total Charge ◽

Colloid Particles

Abstract Background The apoptosis (a cascade of biochemical reactions leading to suicide of damaged biological cells) is blocked in the cancer cells because of impossibility of cytochrome c (cytC) go out from the mitochondria. However, the apoptosis can be started by introducing of exogenous cytC into cytoplasm using colloid particles as a protein carrier due to ability of the cancer cells to phagocytize extracellular particles with submicron size. Results The clay mineral montmorillonite (MM) were used to prepare aqueous suspension of protein/mineral composite particles by electrostatic adsorption of the positively charged cytC globules on the negatively charged MM colloid plates, and then added to colon cancel culture. The results shows out that separately cytC and MM have no effect but the composite cytC-MM particles kill 95% of the cancer cells after 96 h treatment using equine cytC which is 97% structurally identical with the human cytC. To reach this high cytotoxicity we have formulated requirements to: (a) bare colloid particles (electric charge, form and size), (b) conditions for protein adsorption (concentrations, pH, ionic strength), and (c) suspension with the composite particles (positive total charge and optimal concentration). Due to satisfying these requirements we have reached cytotoxicity which is 1/3 higher than the reached by other authors using different artificial particles. The cytotoxicity rapidly increases with concentration of the cytC-MM particles but further it shows tendency to saturation. Methods The optimal pH 6.5 and the 10:3 mg/mg cytC/MM concentration ratio at adsorption were found out by employing computer (protein electrostatics) and physicochemical methods (microelectrophoresis and colloid electrooptics) to prepare cytC-MM suspension. The anticancer capability of cytC-MM nanoplates were investigated using cell culture of metastasizing colon cancer. Conclusion The in vitro experiments with colon cancer cell culture disclose that cytC-MM composite particles have potential for application in anticancer therapy of superficial neoplasms of the skin and the alimentary system (mouth cavity, esophagus, stomach, jejunum and colon). Graphic abstract

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Enzyme-immobilized hydrogels to create hypoxia for in vitro cancer cell culture

Journal of Biotechnology ◽

10.1016/j.jbiotec.2017.03.007 ◽

2017 ◽

Vol 248 ◽

pp. 25-34 ◽

Cited By ~ 8

Author(s):

Camron S. Dawes ◽

Heiko Konig ◽

Chien-Chi Lin

Keyword(s):

Cell Culture ◽

Cancer Cell

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Fabrication of Antheraea pernyi Silk Fibroin-Based Thermoresponsive Hydrogel Nanofibers for Colon Cancer Cell Culture

Polymers ◽

10.3390/polym14010108 ◽

2021 ◽

Vol 14 (1) ◽

pp. 108

Author(s):

Bo-Xiang Wang ◽

Jia Li ◽

De-Hong Cheng ◽

Yan-Hua Lu ◽

Li Liu

Keyword(s):

Cell Culture ◽

Colon Cancer ◽

Cancer Cell ◽

Silk Fibroin ◽

Glycidyl Ether ◽

Polymerization Time ◽

Antheraea Pernyi ◽

Thermoresponsive Hydrogel ◽

Lovo Cells

Antheraea pernyi silk fibroin (ASF)-based nanofibers have wide potential for biomaterial applications due to superior biocompatibility. It is not clear whether the ASF-based nanofibers scaffold can be used as an in vitro cancer cell culture platform. In the current study, we fabricated novel ASF-based thermoresponsive hydrogel nanofibers by aqueous electrospinning for colon cancer (LoVo) cells culture. ASF was reacted with allyl glycidyl ether (AGE) for the preparation of allyl silk fibroin (ASF-AGE), which provided the possibility of copolymerization with allyl monomer. The investigation of ASF-AGE structure by 1H NMR revealed that reactive allyl groups were successfully linked with ASF. ASF-based thermoresponsive hydrogel nanofibers (p (ASF-AGE-NIPAAm)) were successfully manufactured by aqueous electrospinning with the polymerization of ASF and N-isopropylacrylamide (NIPAAm). The p (ASF-AGE-NIPAAm) spinning solution showed good spinnability with the increase of polymerization time, and uniform nanofibers were formed at the polymerization time of 360 min. The obtained hydrogel nanofibers exhibited good thermoresponsive that the LCST was similar with PNIPAAm at about 32 °C, and good degradability in protease XIV PBS solution. In addition, the cytocompatibility of colon cancer (LoVo) cells cultured in hydrogel nanofibers was assessed. It was demonstrated that LoVo cells grown on hydrogel nanofibers showed improved cell adhesion, proliferation, and viability than those on hydrogel. The results suggest that the p (ASF-AGE-NIPAAm) hydrogel nanofibers have potential application in LoVo cells culture in vitro. This study demonstrates the feasibility of fabricating ASF-based nanofibers to culture LoVo cancer cells that can potentially be used as an in vitro cancer cell culture platform.

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A Unique 3D In Vitro Cellular Invasion Assay

CrossRef Listing of Deleted DOIs ◽

10.1177/1087057112449863 ◽

2012 ◽

Vol 17 (8) ◽

pp. 1088-1095 ◽

Cited By ~ 12

Author(s):

Daniela F. Quail ◽

Tamara J. Maciel ◽

Kem Rogers ◽

Lynne M. Postovit

Keyword(s):

Breast Cancer ◽

Cell Culture ◽

Cancer Cell ◽

3D Cell Culture ◽

Breast Cancer Cell Lines ◽

Invasion Assay ◽

Cell Culture Techniques ◽

Culture Techniques ◽

Cellular Invasion

Three-dimensional (3D) cell culture techniques using a bioreactor have been used to co-culture various breast cancer cell lines. Comparisons between 3D co-cultures containing different proportions of breast cancer cell lines have been made with respect to cluster size, cell surface marker distribution, and Ki67 expression. Furthermore, an observed difference in invasion through collagen between co-cultures has been briefly reported. However, these assays have not yet been developed into a quantifiable methodology to assess the effects of drugs and/or microenvironments on cellular invasion. From a cancer perspective, two important aspects of cellular invasion that are often left out of in vitro assays are considerations about the 3D structural heterogeneity of the primary tumor and the ability of cells to migrate in all directions. Accordingly, we have taken advantage of the methodology previously described for 3D cell culture techniques and have developed a 3D invasion assay using cell clusters that can be used to assess the effects of different drugs and treatment conditions on cancer cell invasion. We also describe a novel whole-mount technique that permits fluorescence-based immunolocalization of proteins through the entire tumorsphere, without the need for sectioning. Our assay provides a simple, inexpensive, and physiologically relevant context to study cellular invasion in vitro, in a way that recapitulates an in vivo milieu.

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Perfusion Flow Enhances Viability and Migratory Phenotype in 3D-Cultured Breast Cancer Cells

Annals of Biomedical Engineering ◽

10.1007/s10439-021-02727-w ◽

2021 ◽

Author(s):

Alice Pasini ◽

Joseph Lovecchio ◽

Marilisa Cortesi ◽

Chiara Liverani ◽

Chiara Spadazzi ◽

...

Keyword(s):

Breast Cancer ◽

Cell Culture ◽

Cancer Cells ◽

Cancer Cell ◽

3D Cell Culture ◽

Integrated Approach ◽

Cell Distribution ◽

Conventional Culture ◽

Culture Models

AbstractConventional 2D cell culture, a traditional tool in pre-clinical studies, can hardly be regarded as a representation of a natural cell microenvironment. In this respect, it might result in altered cellular behaviors. To overcome such a limitation, different approaches have been tested to conduct more representative in vitro studies. In particular, the use of 3D cell culture introduces variables, such as cell-cell and cell-extracellular matrix interactions; cell features such as survival, proliferation and migration are consequently influenced. For an example, an enhanced drug resistance and increased invasiveness are shown by cancer cells when cultured in 3D versus 2D conventional culture models. In this setting however, non-uniform cell distribution and biological behaviors appear throughout the scaffold, due to reduced diffusion of oxygen and nutrients. Perfusion in bioreactor systems can be used to improve medium transport. In this line of reasoning, this study proposes a breast cancer cell culture model sustained by an integrated approach that couples a 3D environment and a fluid perfusion. This model improves viability and uniformness of cell distribution, while inducing morphological, functional and molecular cancer cell remodeling.

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Inhibitor of ppGalNAc-T3-mediated O-glycosylation blocks cancer cell invasiveness and lowers FGF23 levels

eLife ◽

10.7554/elife.24051 ◽

2017 ◽

Vol 6 ◽

Cited By ~ 9

Author(s):

Lina Song ◽

Adam D Linstedt

Keyword(s):

Chronic Kidney Disease ◽

Cell Culture ◽

Cancer Cell ◽

Site Specific ◽

A Cell ◽

Cell Invasiveness ◽

Vitro Analysis ◽

Cancer Cell Invasiveness ◽

In Vitro Analysis

Small molecule inhibitors of site-specific O-glycosylation by the polypeptide N-acetylgalactosaminyltransferase (ppGalNAc-T) family are currently unavailable but hold promise as therapeutics, especially if selective against individual ppGalNAc-T isozymes. To identify a compound targeting the ppGalNAc-T3 isozyme, we screened libraries to find compounds that act on a cell-based fluorescence sensor of ppGalNAc-T3 but not on a sensor of ppGalNAc-T2. This identified a hit that subsequent in vitro analysis showed directly binds and inhibits purified ppGalNAc-T3 with no detectable activity against either ppGalNAc-T2 or ppGalNAc-T6. Remarkably, the inhibitor was active in two medically relevant contexts. In cell culture, it opposed increased cancer cell invasiveness driven by upregulated ppGalNAc-T3 suggesting the inhibitor might be anti-metastatic. In cells and mice, it blocked ppGalNAc-T3-mediated glycan-masking of FGF23 thereby increasing its cleavage, a possible treatment of chronic kidney disease. These findings establish a pharmacological approach for the ppGalNAc-transferase family and suggest that targeting specific ppGalNAc-transferases will yield new therapeutics.

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Indoleamine 2,3-Dioxygenase-1 Expression is Changed During Bladder Cancer Cell Invasion

International Journal of Tryptophan Research ◽

10.1177/11786469211065612 ◽

2022 ◽

Vol 15 ◽

pp. 117864692110656

Author(s):

Hellen Joyce Sousa Pereira Santos ◽

Luiz Henrique Gomes Matheus ◽

Aline Silva ◽

Stephanie Vanin Dalmazzo ◽

Andressa Assunção Santos ◽

...

Keyword(s):

Cell Culture ◽

Cancer Cell ◽

Cell Invasion ◽

Bladder Carcinoma ◽

Differentially Expressed ◽

Indoleamine 2,3 Dioxygenase ◽

T24 Cells ◽

Ido1 Expression ◽

Cell Invasiveness

The severity of the bladder carcinoma (BC) is directly linked to cell invasion and metastasis. Indoleamine 2,3-dioxygenase-1 (IDO-1) is an INF-γ-induced immunomodulating enzyme that has been linked to the cancer cell invasiveness. Because IDO1 is variable among the tumors, we analyzed its expression in the BC invasion using BC mice models and cell culture. MB49 cells were orthotopically or ectopically inoculated in C57Bl6 mice to evaluate IDO1 by immunohistochemistry. For in vitro experiments, expression of IDO1 and INF-γ was evaluated in grade-1 (RT4) and in grade-3 (T24) BC cell lines. Invading and non-invading T24 cells were separated using the Matrigel/Transwell system, of which total RNA was extracted immediately or after 2 weeks of subculture. Finally, IDO1 was silenced in T24 cells to verify its role on cell invasiveness. In both animal models, IDO1 was differentially expressed between non-invading and invading cells. In cell culture, T24 cells expressed more IDO1 than RT4 cells, independently of the INF-γ expression. IDO1 was differentially expressed between non-invading and invading T24 cells, a difference that was lost by long-time subculture. IDO1 silencing resulted in diminished cell invasiveness. In conclusion, IDO1 expression is changed during bladder carcinoma invasion, playing an important role in this process.

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