scholarly journals Size-Specific Copper Nanoparticle Cytotoxicity Varies between Human Cell Lines

2021 ◽  
Vol 22 (4) ◽  
pp. 1548
Author(s):  
Ina Na ◽  
David C. Kennedy
Keyword(s):  
Cell Culture ◽  
Cell Lines ◽  
Copper Nanoparticles ◽  
Human Cell ◽  
Time Course ◽  
Culture Media ◽  
Critical Role ◽  
Human Cell Lines ◽  
Cell Culture Media ◽  
Particle Stability

Commercially available copper nanoparticles of three different sizes were tested for cytotoxicity against three human cell lines using four different cytotoxicity assays. This array of data was designed to elucidate trends in particle stability, uptake, and cytotoxicity. The copper nanoparticles are not stable in cell culture media, and rapid changes over the time course of the assays play a critical role in the measured endpoints. Typically, the 40–60 nm particles tested were more cytotoxic than either smaller or larger particles. These particles were also taken up more readily by cells and exhibited different stability dynamics in cell culture media. This provides a good correlation between total cellular uptake of copper and cytotoxicity that may be directly linked to particle stability, though it is unclear why the intermediate-sized particles exhibited these unique properties when compared with both larger and smaller particles.

Using 3D perfusion bioreactor system to studying cell biology of ovarian cancer.

2021 ◽  
Vol 39 (15_suppl) ◽  
pp. e17558-e17558
Author(s):  
Alba Martínez ◽  
Molly Buckley ◽  
Joel Berry ◽  
Rebecca Christian Arend
Keyword(s):  
Ovarian Cancer ◽  
Cell Culture ◽  
Cell Growth ◽  
Cell Lines ◽  
Culture Media ◽  
Tumor Biology ◽  
Perfusion Bioreactor ◽  
Suitable Model ◽  
Cell Culture Media ◽  
Bioreactor System

e17558 Background: Epithelial Ovarian Cancer (EOC) is the most common cause of death among gynecological malignancies. This is a result of the high rate of recurrence and chemo-resistance in EOC patients. Therefore, the development of new therapeutics is crucial. A major factor contributing to this is the lack of therapeutic candidates is lack of translational accuracy in preclinical models. Recently, 3-dimensional (3-D) models have aided in accurately recreating tumor biology. We have developed an EOC 3-D perfused bioreactor system that recapitulates EOC tumor biology and incorporates tumor biomechanical regulation. This model allows for us to more accurately predict the clinical response of new drug candidates, which aids in elimination of ineffective candidates prior to clinical trials. Methods: EOC cell lines (luciferase-taggedSKOV-3 and OVCAR-8) were embedded in a relevant extracellular matrix (ECM) and injected into a perfused, polydimethylsiloxane (PDMS) bioreactor. Microchannels were embedded in matrigel so that the cell culture media with or without chemotherapy could flow through the perfused PDMS to provide nutrient delivery and gas exchange enhancing viability and function of surrounding cells. The bioreactors were connected to a peristaltic pump that allowed for the cell culture media to perfuse over a 7-day period. We monitored cell viability using bioluminescence imaging (BLI), immunohistochemistry (IHC), and lactate dehydrogenase (LDH) release in media. Results: BLI showed a linear increase in SKOV-3 and OVCAR-8 cell growth over 7 days. These results were confirmed by IHC measuring the number of nucleated cells per micron2. Graphical representation of the region of interest (ROI) showed a high correlation between IHC staining of nucleated cells and BLI score. IHC analysis of PAX8 staining was positive and proved that the perfusion bioreactor system maintains EOC biology over time. In addition, our results suggest that the bioreactor is a suitable model for drug preclinical testing in both cell lines as well as in patients’ samples. Conclusions: Our preliminary results using the 3D EOC perfused, PDMS bioreactor model showed increased EOC cell growth overtime, while maintaining original EOC histology. Moreover, our results suggest that this model could provide a novel platform to study therapeutic interventions in EOC. Our ultimate goal is to implement ovarian cancer microenvironment components (e.g. immune cells) into bioreactor system to study different drug treatments to better determine drug candidate’s translational efficacy.

scholarly journals Relative cytotoxicity of complexes of platinum(II) and palladium(II) against pure cell culture Paramecium caudatum and human cell lines A431 and HaCaT

2018 ◽  
Vol 7 (1) ◽  
pp. 28-38 ◽  
Author(s):  
Aleksei Vladimirovich Eremin ◽  
Roman Vladimirovich Suezov ◽  
Polina Sergeevna Grishina ◽  
Alexander Ivanovich Ponyaev ◽  
Nicolay Leonidovich Medvedskiy
Keyword(s):  
Cell Culture ◽  
Cell Lines ◽  
Mtt Assay ◽  
Human Cell ◽  
Human Cells ◽  
Epidermoid Carcinoma ◽  
Binuclear Complexes ◽  
Human Cell Lines ◽  
Paramecium Caudatum ◽  
Pure Cell

The results of cytotoxicity cis-diamine mono- and binuclear complexes of platinum(II) and palladium(II) are presented. The cytotoxicity was investigated by the method of biotesting with Paramecium caudatum and by MTT-assay with human cells: epidermoid carcinoma A431 and minimal transformed aneuploid keratinocytes HaCaT. Cytotoxicity of complexes towards protists is higher than against human cells, however, comparatively, HaCaT is more sensitive than A431 by the treatment all complexes. It is noted that cytotoxicity of palladium(II) complexes is higher than the analogues with platinum(II).

scholarly journals Genotoxicity of TiO2 Nanoparticles in Four Different Human Cell Lines (A549, HEPG2, A172 and SH-SY5Y)

Nanomaterials ◽  
2020 ◽  
Vol 10 (3) ◽  
pp. 412 ◽  
Author(s):  
Fátima Brandão ◽  
Natalia Fernández-Bertólez ◽  
Fernanda Rosário ◽  
Maria João Bessa ◽  
Sónia Fraga ◽  
...  
Keyword(s):  
Cell Lines ◽  
Human Cell ◽  
Culture Media ◽  
Cell Model ◽  
Titanium Dioxide Nanoparticles ◽  
Food Additives ◽  
Toxicity Assessment ◽  
Consumer Products ◽  
Human Cell Lines ◽  
Tio2 Nps

Titanium dioxide nanoparticles (TiO2 NPs) have a wide variety of applications in many consumer products, including as food additives, increasing the concern about the possible hazards that TiO2 NPs may pose to human health. Although most previous studies have focused on the respiratory system, ingestion must also be considered as an important exposure route. Furthermore, after inhalation or ingestion, TiO2 NPs can reach several organs, such as the liver, brain or lungs. Taking this into consideration, the present study focuses on the uptake and potential genotoxicity (micronuclei induction) of TiO2 NPs on four human cell lines of diverse origin: lung cells (A549), liver cells (HepG2), glial cells (A172) and neurons (SH-SY5Y), using flow cytometry methods. Results showed a concentration-, time- and cell-type- dependent increase in TiO2 NPs uptake but no significant induction of micronuclei in any of the tested conditions. Data obtained reinforce the importance of cell model and testing protocols choice for toxicity assessment. However, some questions remain to be answered, namely on the role of cell culture media components on the agglomeration state and mitigation of TiO2 NPs toxic effects.

scholarly journals In vitro cytotoxicity of different dental resin-cements on human cell lines

2021 ◽  
Vol 32 (1) ◽  
Author(s):  
Freya Diemer ◽  
Helmut Stark ◽  
Ernst-Heinrich Helfgen ◽  
Norbert Enkling ◽  
Rainer Probstmeier ◽  
...  
Keyword(s):  
Cell Culture ◽  
Cell Viability ◽  
Cell Lines ◽  
Human Cell ◽  
Zinc Phosphate ◽  
Human Cell Lines ◽  
Cell Type ◽  
Direct Stimulation ◽  
Resin Cements

AbstractAdhesive resin-cements are increasingly used in modern dentistry. Nevertheless, released substances from resin materials have been shown to cause cellular toxic effects. Disc-shaped specimens from 12 different resin cements and one conventional zinc phosphate cement were prepared and used for direct stimulation of five different human cell lines via transwell cell culture system or in an indirect way using conditioned cell culture media. Cytotoxicity was determined using LDH and BCA assays. All tested cements led to a decrease of cell viability but to a distinct extent depending on cell type, luting material, and cytotoxicity assay. In general, cements exhibited a more pronounced cytotoxicity in direct stimulation experiments compared to stimulations using conditioned media. Interestingly, the conventional zinc phosphate cement showed the lowest impact on cell viability. On cellular level, highest cytotoxic effects were detected in osteoblastic cell lines. All resin cements reduced cell viability of human cells with significant differences depending on cell type and cement material. Especially, osteoblastic cells demonstrated a tremendous increase of cytotoxicity after cement exposure. Although the results of this in vitro study cannot be transferred directly to a clinical setting, it shows that eluted substances from resin cements may disturb osteoblastic homeostasis that in turn could lead to conditions favoring peri-implant bone destruction. Thus, the wide use of resin cements in every clinical situation should be scrutinized. A correct use with complete removal of all cement residues and a sufficient polymerization should be given the utmost attention in clinical usage.

scholarly journals Cell Culture Systems To Study Human Herpesvirus 6A/B Chromosomal Integration

2017 ◽  
Vol 91 (14) ◽  
Author(s):  
Annie Gravel ◽  
Isabelle Dubuc ◽  
Nina Wallaschek ◽  
Shella Gilbert-Girard ◽  
Vanessa Collin ◽  
...  
Keyword(s):  
Gene Expression ◽  
Cell Culture ◽  
Cell Lines ◽  
Human Cell ◽  
Histone Deacetylase Inhibitors ◽  
Viral Integration ◽  
Viral Gene ◽  
Human Cell Lines ◽  
Cellular Factors ◽  
Viral Genes

ABSTRACT Human herpesviruses 6A/B (HHV-6A/B) can integrate their viral genomes in the telomeres of human chromosomes. The viral and cellular factors contributing to HHV-6A/B integration remain largely unknown, mostly due to the lack of efficient and reproducible cell culture models to study HHV-6A/B integration. In this study, we characterized the HHV-6A/B integration efficiencies in several human cell lines using two different approaches. First, after a short-term infection (5 h), cells were processed for single-cell cloning and analyzed for chromosomally integrated HHV-6A/B (ciHHV-6A/B). Second, cells were infected with HHV-6A/B and allowed to grow in bulk for 4 weeks or longer and then analyzed for the presence of ciHHV-6. Using quantitative PCR (qPCR), droplet digital PCR, and fluorescent in situ hybridization, we could demonstrate that HHV-6A/B integrated in most human cell lines tested, including telomerase-positive (HeLa, MCF-7, HCT-116, and HEK293T) and telomerase-negative cell lines (U2OS and GM847). Our results also indicate that inhibition of DNA replication, using phosphonoacetic acid, did not affect HHV-6A/B integration. Certain clones harboring ciHHV-6A/B spontaneously express viral genes and proteins. Treatment of cells with phorbol ester or histone deacetylase inhibitors triggered the expression of many viral genes, including U39, U90, and U100, without the production of infectious virus, suggesting that the tested stimuli were not sufficient to trigger full reactivation. In summary, both integration models yielded comparable results and should enable the identification of viral and cellular factors contributing to HHV-6A/B integration and the screening of drugs influencing viral gene expression, as well as the release of infectious HHV-6A/B from the integrated state. IMPORTANCE The analysis and understanding of HHV-6A/B genome integration into host DNA is currently limited due to the lack of reproducible and efficient viral integration systems. In the present study, we describe two quantitative cell culture viral integration systems. These systems can be used to define cellular and viral factors that play a role in HHV-6A/B integration. Furthermore, these systems will allow us to decipher the conditions resulting in virus gene expression and excision of the integrated viral genome resulting in reactivation.

scholarly journals An untargeted metabolomics strategy to measure differences in metabolite uptake and excretion by mammalian cell lines

2020 ◽  
Author(s):  
Marina Wright Muelas ◽  
Ivayla Roberts ◽  
Farah Mughal ◽  
Steve O’Hagan ◽  
Philip J. Day ◽  
...  
Keyword(s):  
Blood Serum ◽  
Cell Lines ◽  
Human Cell ◽  
Time Course ◽  
Phospholipid Bilayer ◽  
Untargeted Metabolomics ◽  
Human Cell Lines ◽  
Biologically Relevant ◽  
Mammalian Cell Lines ◽  
Transporter Expression

AbstractIntroductionIt is widely but erroneously believed that drugs get into cells by passing through the phospholipid bilayer portion of the plasma and other membranes. Much evidence shows, however, that this is not the case, and that drugs cross biomembranes by hitchhiking on transporters for other natural molecules to which these drugs are structurally similar. Untargeted metabolomics can provide a method for determining the differential uptake of such metabolites.ObjectivesBlood serum contains many thousands of molecules and provides a convenient source of biologically relevant metabolites. Our objective was to measure them.MethodsWe develop an untargeted LC-MS/MS method to detect a broad range of compounds present in human serum. We apply this to the analysis of the time course of the uptake and secretion of metabolites in serum by several human cell lines, by analysing changes in the serum that represents the extracellular phase (the ‘exometabolome’ or metabolic footprint).ResultsOur method measures some 4,000-5,000 metabolic features in both ES+ and ES− modes. We show that the metabolic footprints of different cell lines differ greatly from each other.ConclusionOur new, 15-minute untargeted metabolome method allows for the robust and convenient measurement of differences in the uptake of serum compounds by cell lines following incubation in serum, and its relation to differences in transporter expression.

scholarly journals Phosphorylated Osteopontin Secreted from Cancer Cells Induces Cancer Cell Motility

Biomolecules ◽  
2021 ◽  
Vol 11 (9) ◽  
pp. 1323
Author(s):  
Yoshinobu Kariya ◽  
Midori Oyama ◽  
Yukiko Kariya ◽  
Yasuhiro Hashimoto
Keyword(s):  
Cell Culture ◽  
Cell Migration ◽  
Cell Motility ◽  
Cancer Cells ◽  
Cell Lines ◽  
Cancer Cell ◽  
Culture Media ◽  
Cell Culture Media ◽  
Cancer Cell Motility ◽  
H460 Cells

Osteopontin (OPN) plays a pivotal role in cancer cell invasion and metastasis. Although OPN has a large number of phosphorylation sites, the functional significance of OPN phosphorylation in cancer cell motility remains unclear. In this study, we attempted to investigate whether phosphorylated OPN secreted from cancer cells affect cancer cell migration. Quantitative PCR and Western blot analyses revealed that MDA-MB435S, A549, and H460 cells highly expressed OPN, whereas the OPN expression levels in H358, MIAPaca-2, and Panc-1 cells were quite low or were not detected. Compared with the cancer cell lines with a low OPN expression, the high OPN-expressing cancer cell lines displayed a higher cell migration, and the cell migration was suppressed by the anti-OPN antibody. This was confirmed by the OPN overexpression in H358 cancer cells with a low endogenous OPN. Phos-tag ELISA showed that phosphorylated OPN was abundant in the cell culture media of A549 and H460 cells, but not in those of MDA-MB435S cells. Moreover, the A549 and H460 cell culture media, as well as the MDA-MB435S cell culture media with a kinase treatment increased cancer cell motility, both of which were abrogated by phosphatase treatment or anti-OPN antibodies. These results suggest that phosphorylated OPN secreted from cancer cells regulates cancer cell motility.

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