scholarly journals Selectivity of direct plasma treatment and plasma-conditioned media in bone cancer cell lines

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Inès Hamouda ◽  
Cédric Labay ◽  
Uroš Cvelbar ◽  
Maria-Pau Ginebra ◽  
Cristina Canal
Keyword(s):  
Cell Culture ◽  
Plasma Treatment ◽  
Cancer Cell ◽  
Culture Medium ◽  
Culture Media ◽  
Bone Cancer ◽  
Reactive Species ◽  
Conditioned Media ◽  
Cell Culture Medium ◽  
Cell Culture Media

AbstractAtmospheric pressure plasma jets have been shown to impact several cancer cell lines, both in vitro and in vivo. These effects are based on the biochemistry of the reactive oxygen and nitrogen species generated by plasmas in physiological liquids, referred to as plasma-conditioned liquids. Plasma-conditioned media are efficient in the generation of reactive species, inducing selective cancer cell death. However, the concentration of reactive species generated by plasma in the cell culture media of different cell types can be highly variable, complicating the ability to draw precise conclusions due to the differential sensitivity of different cells to reactive species. Here, we compared the effects of direct and indirect plasma treatment on non-malignant bone cells (hOBs and hMSCs) and bone cancer cells (SaOs-2s and MG63s) by treating the cells directly or exposing them to previously treated cell culture medium. Biological effects were correlated with the concentrations of reactive species generated in the liquid. A linear increase in reactive species in the cell culture medium was observed with increased plasma treatment time independent of the volume treated. Values up to 700 µM for H2O2 and 140 µM of NO2− were attained in 2 mL after 15 min of plasma treatment in AdvDMEM cell culture media. Selectivity towards bone cancer cells was observed after both direct and indirect plasma treatments, leading to a decrease in bone cancer cell viability at 72 h to 30% for the longest plasma treatment times while maintaining the survival of non-malignant cells. Therefore, plasma-conditioned media may represent the basis for a potentially novel non-invasive technique for bone cancer therapy.

The biochemical effects of extracellular Zn2+and other metal ions are severely affected by their speciation in cell culture media

Metallomics ◽  
2015 ◽  
Vol 7 (1) ◽  
pp. 102-111 ◽  
Author(s):  
H. Haase ◽  
S. Hebel ◽  
G. Engelhardt ◽  
L. Rink
Keyword(s):  
Cell Culture ◽  
Bovine Serum Albumin ◽  
Culture Medium ◽  
Culture Media ◽  
Zinc Homeostasis ◽  
Cell Culture Medium ◽  
Cell Culture Media ◽  
Biochemical Effects ◽  
Cellular Environment

Differential speciation and lower zinc buffering by less bovine serum albumin (BSA) in cell culture medium lead to altered zinc homeostasis compared to the cellular environmentin vivo.

scholarly journals Serum-free medium increases the replication rate of the avian coronavirus infectious bronchitis virus in chicken embryo kidney cells

2021 ◽  
Author(s):  
José A. Quinteros ◽  
Glenn F. Browning ◽  
Amir H. Noormohammadi ◽  
Mark A. Stevenson ◽  
Mauricio J. C. Coppo ◽  
...  
Keyword(s):  
Cell Culture ◽  
Cell Cultures ◽  
Infectious Bronchitis Virus ◽  
Chicken Embryo ◽  
Culture Medium ◽  
Culture Media ◽  
Cell Culture Medium ◽  
Cell Culture Media ◽  
Infectious Bronchitis ◽  
Porcine Epidemic Diarrhoea Virus

AbstractInfectious bronchitis virus (IBV), an avian coronavirus, can be isolated and cultured in tracheal organ cultures (TOCs), embryonated eggs and cell cultures. TOCs and embryonated eggs are commonly used for viral isolation but use of these is laborious and expensive. Cell cultures have been used only with IBV strains that have previously been adapted to grow under laboratory conditions, and not for primary isolation. Previous studies using the coronavirus porcine epidemic diarrhoea virus (PEDV) have suggested that foetal bovine serum (FBS), a common component of cell culture media, can inhibit the adsorption of coronaviruses onto the host cell membrane receptors. In the present study, the replication of IBV in primary chicken embryo kidney (CEK) cell cultures and the Leghorn hepatocellular carcinoma (LMH) cell line was examined using two different cell culture media, one containing FBS and the other containing yeast extract (YE). A reverse transcriptase quantitative polymerase chain reaction (RT-qPCR) assay was used to quantify viral RNA copies in cell lysates. The highest concentrations of viral genomes were observed when the cell culture medium did not contain FBS. Examination of the infectivity of virus grown in CEK cell cultures was examined by titration in embryonated chicken eggs, demonstrating that the cell lysate from CEK cell cultures in medium without FBS contained a higher median embryo infectious dose (EID50) than that from CEK cell cultures in medium containing FBS. These results suggest that improved replication of IBV in cell cultures can be achieved by the omission of FBS from the cell culture medium. This may enhance the potential for production of vaccines in cell culture and facilitate the isolation of emergent IBV strains in cell cultures.

scholarly journals The Cell Culture Medium Affects Growth, Phenotype Expression and the Response to Selenium Cytotoxicity in A549 and HepG2 Cells

Antioxidants ◽  
2019 ◽  
Vol 8 (5) ◽  
pp. 130 ◽  
Author(s):  
Lisa Arodin Selenius ◽  
Marita Wallenberg Lundgren ◽  
Rim Jawad ◽  
Olof Danielsson ◽  
Mikael Björnstedt
Keyword(s):  
Cell Culture ◽  
Cell Growth ◽  
Culture Medium ◽  
Culture Media ◽  
A549 Cells ◽  
Culture Conditions ◽  
Cell Culture Medium ◽  
Cell Culture Media ◽  
Toxic Response ◽  
Selenium Compounds

Selenium compounds influence cell growth and are highly interesting candidate compounds for cancer chemotherapy. Over decades an extensive number of publications have reported highly efficient growth inhibitory effects with a number of suggested mechanisms f especially for redox-active selenium compounds. However, the studies are difficult to compare due to a high degree of variations in half-maximal inhibitor concentration (IC50) dependent on cultivation conditions and methods to assess cell viability. Among other factors, the variability in culture conditions may affect the experimental outcome. To address this, we have compared the maintenance effects of four commonly used cell culture media on two cell lines, A549 and HepG2, evaluated by the toxic response to selenite and seleno-methylselenocysteine, cell growth and redox homeostasis. We found that the composition of the cell culture media greatly affected cell growth and sensitivity to selenium cytotoxicity. We also provided evidence for change of phenotype in A549 cells when maintained under different culture conditions, demonstrated by changes in cytokeratin 18 (CK18) and vimentin expression. In conclusion, our results have shown the importance of defining the cell culture medium used when comparing results from different studies.

scholarly journals The landscape of metabolic pathway dependencies in cancer cell lines

2021 ◽  
Vol 17 (4) ◽  
pp. e1008942
Author(s):  
James H. Joly ◽  
Brandon T. L. Chew ◽  
Nicholas A. Graham
Keyword(s):  
Cell Culture ◽  
Cancer Cells ◽  
Cell Lines ◽  
Cancer Cell ◽  
Metabolic Pathway ◽  
Culture Medium ◽  
Cancer Cell Lines ◽  
Metabolic Reprogramming ◽  
Cell Culture Medium ◽  
Loss Of Function

The metabolic reprogramming of cancer cells creates metabolic vulnerabilities that can be therapeutically targeted. However, our understanding of metabolic dependencies and the pathway crosstalk that creates these vulnerabilities in cancer cells remains incomplete. Here, by integrating gene expression data with genetic loss-of-function and pharmacological screening data from hundreds of cancer cell lines, we identified metabolic vulnerabilities at the level of pathways rather than individual genes. This approach revealed that metabolic pathway dependencies are highly context-specific such that cancer cells are vulnerable to inhibition of one metabolic pathway only when activity of another metabolic pathway is altered. Notably, we also found that the no single metabolic pathway was universally essential, suggesting that cancer cells are not invariably dependent on any metabolic pathway. In addition, we confirmed that cell culture medium is a major confounding factor for the analysis of metabolic pathway vulnerabilities. Nevertheless, we found robust associations between metabolic pathway activity and sensitivity to clinically approved drugs that were independent of cell culture medium. Lastly, we used parallel integration of pharmacological and genetic dependency data to confidently identify metabolic pathway vulnerabilities. Taken together, this study serves as a comprehensive characterization of the landscape of metabolic pathway vulnerabilities in cancer cell lines.

scholarly journals The Effect of Oxygen and Micronutrient Composition of Cell Growth Media on Cancer Cell Bioenergetics and Mitochondrial Networks

Biomolecules ◽  
2021 ◽  
Vol 11 (8) ◽  
pp. 1177
Author(s):  
Fereshteh Moradi ◽  
Christopher Moffatt ◽  
Jeffrey A. Stuart
Keyword(s):  
Cell Culture ◽  
Energy Metabolism ◽  
Cancer Cell ◽  
Network Structure ◽  
Culture Media ◽  
Growth Media ◽  
Media Composition ◽  
Cell Culture Media ◽  
Lactate Dehydrogenase Activity ◽  
Huh7 Cells

Cancer cell culture is routinely performed under superphysiologic O2 levels and in media such as Dulbecco’s Modified Eagle Medium (DMEM) with nutrient composition dissimilar to mammalian extracellular fluid. Recently developed cell culture media (e.g., Plasmax, Human Plasma-Like Medium (HPLM)), which are modeled on the metabolite composition of human blood plasma, have been shown to shift key cellular activities in several cancer cell lines. Similar effects have been reported with respect to O2 levels in cell culture. Given these observations, we investigated how media composition and O2 levels affect cellular energy metabolism and mitochondria network structure in MCF7, SaOS2, LNCaP, and Huh7 cells. Cells were cultured in physiologic (5%) or standard (18%) O2 levels, and in physiologic (Plasmax) or standard cell culture media (DMEM). We show that both O2 levels and media composition significantly affect mitochondrial abundance and network structure, concomitantly with changes in cellular bioenergetics. Extracellular acidification rate (ECAR), a proxy for glycolytic activity, was generally higher in cells cultured in DMEM while oxygen consumption rates (OCR) were lower. This effect of media on energy metabolism is an important consideration for the study of cancer drugs that target aspects of energy metabolism, including lactate dehydrogenase activity.

scholarly journals The landscape of metabolic pathway dependencies in cancer cell lines

2020 ◽  
Author(s):  
James H. Joly ◽  
Brandon T.L. Chew ◽  
Nicholas A. Graham
Keyword(s):  
Cell Culture ◽  
Cancer Cells ◽  
Cell Lines ◽  
Cancer Cell ◽  
Metabolic Pathway ◽  
Culture Medium ◽  
Cancer Cell Lines ◽  
Metabolic Reprogramming ◽  
Cell Culture Medium ◽  
Loss Of Function

AbstractThe metabolic reprogramming of cancer cells creates metabolic vulnerabilities that can be therapeutically targeted. However, our understanding of metabolic dependencies and the pathway crosstalk that creates these vulnerabilities in cancer cells remains incomplete. Here, by integrating gene expression data with genetic loss-of-function and pharmacological screening data from hundreds of cancer cell lines, we identified metabolic vulnerabilities at the level of pathways rather than individual genes. This approach revealed that metabolic pathway dependencies are highly context-specific such cancer cells are vulnerable to inhibition of one metabolic pathway only when activity of another metabolic pathway is altered. Notably, we also found that the no single metabolic pathway was universally essential, suggesting that cancer cells are not invariably dependent on any metabolic pathway. In addition, we confirmed that cell culture medium is a major confounding factor for the analysis of metabolic pathway vulnerabilities. Nevertheless, we found robust associations between metabolic pathway activity and sensitivity to clinically approved drugs that were independent of cell culture medium. Lastly, we used parallel integration of pharmacological and genetic dependency data to confidently identify metabolic pathway vulnerabilities. Taken together, this study serves as a comprehensive characterization of the landscape of metabolic pathway vulnerabilities in cancer cell lines.

scholarly journals Corynebacterium diphtheriae Proteome Adaptation to Cell Culture Medium and Serum

Proteomes ◽  
2021 ◽  
Vol 9 (1) ◽  
pp. 14
Author(s):  
Jens Möller ◽  
Fatemeh Nosratabadi ◽  
Luca Musella ◽  
Jörg Hofmann ◽  
Andreas Burkovski
Keyword(s):  
Cell Culture ◽  
Growth Medium ◽  
Culture Medium ◽  
Culture Media ◽  
Calf Serum ◽  
Corynebacterium Diphtheriae ◽  
Protein Quantification ◽  
Cell Culture Medium ◽  
Animal Testing ◽  
Additional Advantage

Host-pathogen interactions are often studied in vitro using primary or immortal cell lines. This set-up avoids ethical problems of animal testing and has the additional advantage of lower costs. However, the influence of cell culture media on bacterial growth and metabolism is not considered or investigated in most cases. To address this question growth and proteome adaptation of Corynebacterium diphtheriae strain ISS3319 were investigated in this study. Bacteria were cultured in standard growth medium, cell culture medium, and fetal calf serum. Mass spectrometric analyses and label-free protein quantification hint at an increased bacterial pathogenicity when grown in cell culture medium as well as an influence of the growth medium on the cell envelope.

scholarly journals The Predictive Value of the Pentraxin 3 Concentration in Cumulus Cell Culture Media for the Embryo Implantation

2021 ◽  
pp. 1-7
Author(s):  
Tulay Irez ◽  
Yavuz Sahin ◽  
Eduard Malik ◽  
Onur Guralp
Keyword(s):  
Cell Culture ◽  
Culture Medium ◽  
Culture Media ◽  
Embryo Implantation ◽  
Cumulus Cell ◽  
Culture Fluid ◽  
Cumulus Cells ◽  
Pentraxin 3 ◽  
Cell Culture Media ◽  
Morphological Criteria

OBJECTIVE: Many studies on the interrelation of cumulus cells and oocytes, and research on cumulus protein factors continue. This study aims to investigate the relationship of Pentraxin 3 level with embryo implantation in the cumulus culture fluid. STUDY DESIGN: A total of 31 women with idiopathic infertility who underwent intracytoplasmic sperm injection treatment were prospectively evaluated. Cell suspensions containing 5 million/mL cumulus cells were obtained post-hyase and incubated for 24 hours in a culture medium. A possible association between the culture media Pentraxin 3 concentrations and embryo implantation was analyzed. RESULTS: The cumulus cell culture media Pentraxin 3 concentrations were significantly higher in the pregnant group compared to the non-pregnant group (98.9 ng/mL vs 53.2 ng/mL, respectively, p=0.005). There was a significant positive correlation between the culture media Pentraxin 3 concentrations and embryo implantation (r=0.500, p=0.005). The culture media Pentraxin 3 concentration was a significant predictor for successful embryo implantation (AUC=0.845, p=0.006). A cut-off value of 64.25 ng/mL had an 86% sensitivity and 80% specificity to predict embryo implantation. There was no pregnancy under the cut-off value of 60 ng/mL, whereby seven women had good quality grade 1 oocytes according to the traditional morphological criteria. CONCLUSION: The cumulus cell culture medium Pentraxin 3 concentration was predictive for successful embryo implantation. Low Pentraxin 3 levels (<60 ng/mL were associated with failure of conception.

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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