The Development and Evaluation of Brain and Heart Cell Lines from a Marine Fish for Use in Xenobiotic-Induced Cytotoxicity Testing

2021 ◽  
pp. 026119292110525
Author(s):  
Anaguiven Avalos-Soriano ◽  
Alejandra García-Gasca ◽  
Beatriz Yáñez-Rivera
Keyword(s):  
Cell Lines ◽  
Molecular Mechanisms ◽  
Fetal Bovine Serum ◽  
Cellular Level ◽  
Heart Cell ◽  
Mechanisms Of Toxicity ◽  
Toxicological Data ◽  
Fetal Bovine ◽  
Fish Cells ◽  
The Brain

Two cell lines derived from the brain and heart of a Pacific white snook specimen ( Centropomus viridis) were developed and evaluated in terms of their responsiveness to glyphosate-induced cytotoxicity. The cells were grown in Leibovitz-15 (L-15) medium supplemented with 10% fetal bovine serum (FBS) and were passaged 36 times. Growth was tested at different concentrations of FBS (5, 10 and 20%) at 27°C. The cell lines were cryopreserved at different passages and were successfully thawed, with a survival rate greater than 80% without detectable contamination. At passage 36, the cells were used to assess the deleterious effects of glyphosate, and cell proliferation was measured by direct counting and with the MTT assay. Similar LC50 values were obtained with both methods. Although the principles behind these two assessment methods differ, our results show that both are suitable for evaluating glyphosate toxicity. In addition, heart- and brain-derived cells showed similar sensitivity, suggesting that the same mode of action might be responsible for the toxicity of glyphosate at the cellular level. The newly developed Pacific white snook brain and heart cell lines could be useful to investigate cellular and molecular mechanisms of toxicity, satisfying the need to reduce the use of animals in experiments. Glyphosate-related toxicological data obtained in the present study will allow us to continue investigating the effects of this herbicide directly on brain and heart fish cells since similar studies have only been carried out on either live organisms or on human cell lines such as neuroblastoma, which are immortalised by oncogenes or similar.

scholarly journals Various factors affect lipopolysaccharide sensitization in cell cultures

BioTechniques ◽  
2020 ◽  
Vol 69 (2) ◽  
pp. 126-132
Author(s):  
Nan Yang ◽  
Don D Sin ◽  
Delbert R Dorscheid
Keyword(s):  
Cell Lines ◽  
Cell Cultures ◽  
Bovine Serum ◽  
Fetal Bovine Serum ◽  
Signal Loss ◽  
The Media ◽  
Fetal Bovine

Commercially available lipopolysaccharide (LPS) is commonly used in research. Although protocols for its use are well established, we experienced a loss of LPS responsiveness in our cell cultures despite no obvious experimental changes. Our cell lines were stimulated with LPS and the media quantified for LPS responsiveness via an IL-8 ELISA. We discovered that the major cause of signal loss was differences in fetal bovine serum (FBS) formulation and concentration. One FBS formulation was notably better at eliciting an IL-8 signal than the second FBS, and 10% FBS in media was better at inducing LPS responsiveness than lower concentrations. We urge researchers to be aware of inherent variations in seemingly commonplace reagents as they may be unexpected sources of inconsistencies.

scholarly journals Growth properties and alloantigenic expression of murine lymphoblastoid cell lines.

1975 ◽  
Vol 142 (2) ◽  
pp. 378-390 ◽  
Author(s):  
R K Zwerner ◽  
R T Acton
Keyword(s):  
Cell Line ◽  
Cell Lines ◽  
Fetal Bovine Serum ◽  
Horse Serum ◽  
Culture Conditions ◽  
Lymphoblastoid Cell Lines ◽  
Stationary Growth ◽  
Growth Properties ◽  
Fetal Bovine ◽  
Logarithmic Growth

Murine lymphoblastoid cell lines were evaluated for their expression of Thy-1 and thymus leukemia (TL) differentiation alloantigens. Two culture conditions were shown to affect this expression. Cells grown in fetal bovine serum (FBS)-enriched medium expressed up to 15 times the amount of TL as cells grown in horse serum (HS)-enriched medium. Thy-1 expression was less affected by the type of serum used for culture. The phase of growth when the cells were harvested, was demonstrated to affect the expression of Thy-1. The expression of Thy-1.2 for one cell line examined, L-251A, during logarithmic growth was threefold greater than cells collected during either lag or stationary growth. When culture conditions were standardized a ranking of the amount of Thy-1 and TL expressed by several cell lines was made. All cell lines, except one, L-1210, expressed Thy-1. There was a 450-fold difference in the expression of Thy-1 between the cell lines evaluated. Seven cell lines expressed TL-1,2,3 with a ninefold difference in the amount of expression. The L-251A cell line was cultured in a 14 liter fermentor for a 26 day period. During this time TL and Thy-1 expression did not vary significantly, demonstrating that lymphoblastoid cell lines can be cultured on a continuous basis and will continue to express their surface alloantigens.

scholarly journals Evaluation of the major royal jelly proteins as an alternative to fetal bovine serum in culturing human cell lines

2016 ◽  
Vol 17 (6) ◽  
pp. 476-483 ◽  
Author(s):  
Di Chen ◽  
Xiao-xuan Xin ◽  
Hao-cheng Qian ◽  
Zhang-yin Yu ◽  
Li-rong Shen
Keyword(s):  
Cell Lines ◽  
Human Cell ◽  
Bovine Serum ◽  
Royal Jelly ◽  
Fetal Bovine Serum ◽  
Human Cell Lines ◽  
Fetal Bovine

Effect of fetal bovine serum on the growth and survival of insect cell cultures

1970 ◽  
Vol 48 (3) ◽  
pp. 427-432 ◽  
Author(s):  
S. S. Sohi ◽  
Cheryl Smith
Keyword(s):  
Adverse Effect ◽  
Cell Lines ◽  
Insect Cell ◽  
Bovine Serum ◽  
Fetal Bovine Serum ◽  
Maximum Growth ◽  
Growth And Survival ◽  
Fetal Bovine ◽  
High Concentrations ◽  
Insect Cell Lines

The effect of different concentrations of fetal bovine serum (FBS) on the growth and survival of three insect cell lines—Aedes aegypti, Antheraea eucalypti, and Bombyx mori—was studied in a series of 10 experiments. Cell viability was low and no growth occurred in any of the cell lines when FBS was omitted from the medium. Maximum growth of A. aegypti cells was obtained with 10% FBS. There was no further increase in this growth when FBS was increased to 20 and 30%, and neither did the increased concentrations have any appreciable adverse effect on the growth or survival of these cells. Maximum growth of A. eucalypti and B. mori cells was obtained in 5% FBS; the growth of these cells was significantly less in 20 and 30% FBS. Viability of A. eucalypti cells was quite low in 20 and 30% FBS. There was, however, no adverse effect on the viability of B. mori cells at these high concentrations.

scholarly journals Neuroprotective effects of physical exercise: Implications in health and disease

2021 ◽  
Vol 68 (3) ◽  
pp. 383-389
Author(s):  
Sebastian Romeo Pintilie ◽  
◽  
Alice D. Condrat ◽  
Adriana Fodor ◽  
Adela-Viviana Sitar-Tăut ◽  
...  
Keyword(s):  
Physical Activity ◽  
Physical Exercise ◽  
Molecular Mechanisms ◽  
Cellular Level ◽  
Neuroprotective Effects ◽  
Chemical Effects ◽  
Physical Exercises ◽  
Health And Disease ◽  
Molecular Aspects ◽  
The Brain

Physical exercises have long been linked to numerous health improvements, ranging from cardiovascular to psychiatric. In this review, we take a closer look on its anatomical, physiological and chemical effects on the brain. Starting from the clinical to the cellular level, we will analyze the neurogenesis, anti-inflammatory effects on Brain-Blood Barrier and synaptic plasticity, outlining known molecular aspects that are influenced by physical activity, such as: gene expression, changes of growth factors and neurotransmitter levels and means of reverting molecular mechanisms of ageing. The brain derived neurotrophic factor (BDNF) is one of the central molecules that links the physical exercise to neurogenesis, neuroprotection, cognitive functions, dendritic growth, memory formation and many more. We indicate the correlation between physical activity and mental health in diseases like depression, Alzheimer’s dementia and Parkinson’s disease.

Gamma-Tocotrienol and Simvastatin Synergistically Induce Cytotoxicity In Leukemia Cell Lines, K-562 and HL-60

Blood ◽  
2013 ◽  
Vol 122 (21) ◽  
pp. 4927-4927 ◽  
Author(s):  
Ko Maung ◽  
Victoria Palau ◽  
Janet Lightner ◽  
Marianne Brannon ◽  
Koyamangalath Krishnan
Keyword(s):  
Cell Lines ◽  
Fetal Bovine Serum ◽  
Cholesterol Biosynthesis ◽  
Leukemia Cell ◽  
Leukemia Cells ◽  
Hmg Coa Reductase ◽  
Leukemia Cell Lines ◽  
Fetal Bovine ◽  
Coa Reductase ◽  
Gamma Tocotrienol

Abstract Introduction Statins and tocotrienols modulate the cholesterol biosynthesis pathway by inhibiting the 3-hydroxy-3- methylglutaryl coenzyme A (HMG-CoA) reductase. Tocotrienols modulate HMG-CoA reductase by post-transcriptional downregulation. In addition, tocotrienols contain a farnesol moiety in its side-chain that triggers degradation of HMG-CoA reductase. These effects lead to suppression of cell proliferation, cell cycle arrest, and apoptosis. Several studies have shown that statins have suppressive effects in in vitro experiments on acute myelocytic leukemia cell lines. Since both statins and gamma-tocotrienol are associated with decreased cholesterol biosynthesis, we hypothesized that if the cytotoxicity of these drugs on cancer cells is related to impaired biosynthesis of cholesterol, combination of them could synergize in cytotoxicity on leukemic cells. Materials and Methods K-562 and HL-60 leukemia cells were grown in Iscove's Modified Dulbecco's medium with penicillin/streptomycin, 10% fetal bovine serum and 20% fetal bovine serum added respectively. K-562 and HL-60 leukemia cells were seeded in 96 well plates, grown overnight, and treated for 24, 48 and 72 hours with simvastatin in concentrations of 1,2,4, and5 µM; gamma-tocotrienol in concentrations of 20, 40, and 80 µM; and a combination of the two drugs in the same concentrations. For 24 hour dose, cells were seeded at a density of 5000/well and for 48 and 72 hour doses, at  3500/well. Following the treatment, MTS/PMS reagent (Promega, Madison, WI), [3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt] mixed with and an electron coupling reagent (phenazine methosulfate), was added at 40 µg/well and incubated at 37°C for 2 hours. We measured the solubilized formazan crystals at 450 nm as an indicator of cytotoxicity. The presence of ATP  as an indicator of cell viability was measured with the CellTiter Glo®  assay (Promega). Cells were again seeded, grown overnight, and dosed as indicated above.  Following treatment, the assay was conducted as specified by the manufacturer. Results Both simvastatin and gamma-tocotrienol induce cytotoxicity in K-562 and HL-60 cell lines by the MTS and Cell Titer Glo Assays.  The IC50 at 72 hour incubation are as follows 1) HL-60: simvastatin - 16.543 uM, gamma tocotrienol - 36.297 uM;  2) K-562: simvastatin - 5.235 uM, gamma tocotrienol - 34.947 uM.  We used CompuSyn to calculate the IC50.  When combined, simvastatin and gamma-tocotrienol exhibit synergy at lower concentrations when examined by isobologram analysis. Conclusion Gamma-tocotrienol, an isoform of vitamin E, and simvastatin, a cholesterol lowering drug exhibit synergy in induction of cytotoxicity in K-562 and HL-60 leukemia cell lines. Rescue experiments and mechanistic pathway analysis are being explored to confirm these observations. Disclosures: No relevant conflicts of interest to declare.

Characterization of osteoblastic properties of 7F2 and UMR-106 cultures after acclimation to reduced levels of fetal bovine serum

2008 ◽  
Vol 86 (7) ◽  
pp. 403-415 ◽  
Author(s):  
S. Ganguly ◽  
L.A. Ashley ◽  
C.M. Pendleton ◽  
R.D. Grey ◽  
G.C. Howard ◽  
...  
Keyword(s):  
Cell Lines ◽  
Bovine Serum ◽  
Fetal Bovine Serum ◽  
Phenol Red ◽  
Ph Indicator ◽  
Fetal Bovine ◽  
Experimental Findings ◽  
Umr 106 ◽  
Culture Protocol

Estrogen plays an important role in skeletal physiology by maintaining a remodeling balance between the activity of osteoblasts and osteoclasts. In an attempt to decipher the mechanism through which estrogen elicits its action on osteoblasts, experimentation necessitated the development of a culturing environment reduced in estrogenic compounds. The selected medium (OPTI-MEM) is enriched to sustain cultures under reduced fetal bovine serum (FBS) conditions and is devoid of the pH indicator phenol red, a suspected estrogenic agent. This protocol reduced the concentration of FBS supplementation to 0% through successive 24 h incubations with diminishing amounts of total FBS (1%, 0.1%, and 0%). The protocol does not appear to alter the viability, cell morphology, or osteoblast-like phenotype of 7F2 and UMR-106 cell lines when compared with control cells grown in various concentrations of FBS. Although the rate of mitotic divisions declined, the 7F2 and UMR-106 cultures continued to express osteoblast-specific markers and exhibited estrogen responsiveness. These experimental findings demonstrate that the culture protocol developed did not alter the osteoblast nature of the cell lines and provides a model system to study estrogen’s antiresorptive role on skeletal turnover.

scholarly journals Putative Role of MicroRNA-Regulated Pathways in Comorbid Neurological and Cardiovascular Disorders

2009 ◽  
Vol 2009 ◽  
pp. 1-5 ◽  
Author(s):  
Sébastien S. Hébert
Keyword(s):  
Mirna Expression ◽  
Molecular Mechanisms ◽  
Expression Profiles ◽  
Heart Cell ◽  
Cardiovascular Disorders ◽  
Increased Risk ◽  
Mirna Expression Profiles ◽  
And Function ◽  
The Brain

Background. The conserved noncoding microRNAs (miRNAs) that function to regulate gene expression are essential for the development and function of the brain and heart. Changes in miRNA expression profiles are associated with an increased risk for developing neurodegenerative disorders as well as heart failure. Here, the hypothesis of how miRNA-regulated pathways could contribute to comorbid neurological and cardiovascular disorders will be discussed. Presentation. Changes in miRNA expression occurring in the brain and heart could have an impact on coexisting neurological and cardiovascular characteristics by (1) modulating organ function, (2) accentuating cellular stress, and (3) impinging on neuronal and/or heart cell survival. Testing. Evaluation of miRNA expression profiles in the brain and heart tissues from individuals with comorbid neurodegenerative and cardiovascular disorders will be of great importance and relevance. Implications. Careful experimental design will shed light to the deeper understanding of the molecular mechanisms tying up those different but yet somehow connected diseases.

scholarly journals Alterations in Cell Motility, Proliferation, and Metabolism in Novel Models of Acquired Temozolomide Resistant Glioblastoma

2017 ◽  
Author(s):  
DM Tiek ◽  
JD Rone ◽  
GT Graham ◽  
EL Pannkuk ◽  
BR Haddad ◽  
...  
Keyword(s):  
Cell Lines ◽  
Molecular Mechanisms ◽  
Rapid Development ◽  
Treatment Options ◽  
Standard Of Care ◽  
Model Systems ◽  
Effective Strategies ◽  
Development Of Resistance ◽  
The Brain ◽  
Line Standard

AbstractGlioblastoma (GBM) is an aggressive and incurable tumor of the brain with limited treatment options. Current first-line standard of care is the DNA alkylating agent temozolomide (TMZ), but this treatment strategy adds only ~4 months to median survival due to the rapid development of resistance. While some mechanisms of TMZ resistance have been identified, they are not fully understood. There are few effective strategies to manage therapy resistant GBM, and we lack diverse preclinical models of acquired TMZ resistance in which to test therapeutic strategies on TMZ resistant GBM. In this study, we create and characterize two new GBM cell lines resistant to TMZ, based on the 8MGBA and 42MGBA cell lines. Analysis of the TMZ resistant (TMZres) variants in conjunction with their parental, sensitive cell lines shows that acquisition of TMZ resistance is accompanied by broad phenotypic changes, including increased proliferation, migration, chromosomal aberrations and secretion of cytosolic lipids. Importantly, each TMZ resistant model captures a different facet of the “go” (8MGBA-TMZres) or “grow” (42MGBA-TMZres) hypothesis of GBM behavior. These model systems will be important additions to the available tools for investigators seeking to define molecular mechanisms of acquired TMZ resistance.

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