Static Magnetic Stimulation Induces Changes in the Oxidative Status and Cell Viability Parameters in a Primary Culture Model of Astrocytes

2021 ◽  
Author(s):  
Caroline Crespo da Costa ◽  
Léo Anderson Meira Martins ◽  
André Peres Koth ◽  
Jéssica Marques Obelar Ramos ◽  
Fátima Theresinha Costa Rodrigues Guma ◽  
...  
Keyword(s):  
Cell Viability ◽  
Primary Culture ◽  
Magnetic Stimulation ◽  
Oxidative Status ◽  
Culture Model

scholarly journals Zinc, Zinc Transporters, and Cadmium Cytotoxicity in a Cell Culture Model of Human Urothelium

Toxics ◽  
2021 ◽  
Vol 9 (5) ◽  
pp. 94
Author(s):  
Soisungwan Satarug ◽  
Scott H. Garrett ◽  
Seema Somji ◽  
Mary Ann Sens ◽  
Donald A. Sens
Keyword(s):  
Dna Methylation ◽  
Cell Culture ◽  
Cell Viability ◽  
Zinc Transporters ◽  
Cell Culture Model ◽  
Induced Expression ◽  
Glutathione Biosynthesis ◽  
Culture Model ◽  
A Cell ◽  
Cd Exposure

We explored the potential role of zinc (Zn) and zinc transporters in protection against cytotoxicity of cadmium (Cd) in a cell culture model of human urothelium, named UROtsa. We used real-time qRT-PCR to quantify transcript levels of 19 Zn transporters of the Zrt-/Irt-like protein (ZIP) and ZnT gene families that were expressed in UROtsa cells and were altered by Cd exposure. Cd as low as 0.1 µM induced expression of ZnT1, known to mediate efflux of Zn and Cd. Loss of cell viability by 57% was seen 24 h after exposure to 2.5 µM Cd. Exposure to 2.5 µM Cd together with 10–50 µM Zn prevented loss of cell viability by 66%. Pretreatment of the UROtsa cells with an inhibitor of glutathione biosynthesis (buthionine sulfoximine) diminished ZnT1 induction by Cd with a resultant increase in sensitivity to Cd cytotoxicity. Conversely, pretreatment of UROtsa cells with an inhibitor of DNA methylation, 5-aza-2’-deoxycytidine (aza-dC) did not change the extent of ZnT1 induction by Cd. The induced expression of ZnT1 that remained impervious in cells treated with aza-dC coincided with resistance to Cd cytotoxicity. Therefore, expression of ZnT1 efflux transporter and Cd toxicity in UROtsa cells could be modulated, in part, by DNA methylation and glutathione biosynthesis. Induced expression of ZnT1 may be a viable mechanistic approach to mitigating cytotoxicity of Cd.

MO1048PHYTOTHERAPY ON RENAL HYPOXIA: THE PREVENTIVE EFFECTS OF HYDROXYTYROSOL - PRELIMINARY IN VITRO RESULTS

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
Terézia Kamasová ◽  
Ana Sofia Abreu ◽  
Fátima Paiva-Martins ◽  
Luís Belo ◽  
Alice Santos-Silva ◽  
...  
Keyword(s):  
Cell Viability ◽  
Cell Line ◽  
Therapeutic Potential ◽  
Quantitative Polymerase Chain Reaction ◽  
Oxidative Status ◽  
Hypoxic Conditions ◽  
Thiazolyl Blue Tetrazolium Bromide ◽  
Anti Inflammatory ◽  
Preventive Effects

Abstract Background and Aims Renal hypoxia plays a key role in the pathophysiology of acute kidney injury and in the progression of chronic kidney disease, potentiating other important risk factors for renal disease, such as oxidative stress, renal fibrosis, and inflammation. Hydroxytyrosol (HT) is a phenolic compound extracted from olives and olive-derived products, that has been shown to detain potent in vitro antioxidant and anti-inflammatory activity. The aim of this study was to evaluate the preventive therapeutic potential of HT on a cellular model of renal hypoxia. Method A cell line of normal adult proximal tubular epithelium (HK-2 cell line) was used to determine the effects of the chemical induction of hypoxia with cobalt chloride (CoCl2), as well as the preventive potential of HT on the elicited effects. For this purpose, HK-2 cells were exposed for 24 h to 254 µM CoCl2, to mimic the hypoxic conditions, or pre-incubated for 1 h with 5 µM HT and further exposed to the CoCl2 for 24 h more. Cell viability was assessed by the thiazolyl blue tetrazolium bromide reduction assay. Oxidative status was evaluated by the measurement of reactive oxygen and nitrogen species (ROS and RNS) and reduced glutathione (GSH) levels, by using standardized fluorometric and colorimetric assays. The expression of several genes related to the hypoxic, inflammatory, and fibrotic responses was determined by quantitative polymerase chain reaction (PCR). Results CoCl2-exposed HK-2 cells (hypoxic conditions) showed a significant decrease in cell viability (p < 0.0001 vs. control), and a disruption of the oxidative status, characterized by an increase of ROS and RNS production of about 6-fold over control cells (p < 0.0001) and a decrease in GSH intracellular levels of nearly 50 % (p < 0.05). Although the pre-exposure to HT showed no significant effects on the loss of cell viability elicited by CoCl2, the presence of HT prior to induction of hypoxia reduced the generation of ROS and RNS (p < 0.05 for HT + CoCl2 vs. CoCl2) and prevented the GSH depletion (GSH levels for HT + CoCl2 were similar to those of control) elicited by CoCl2. When compared to control cells, CoCl2-exposed HK-2 cells also showed increased expression of genes related to hypoxia (HIF1A, p < 0.05; GAPDH, p < 0.0001), as well as of modulators of inflammation (IL6, p < 0.0001) and fibrosis (TGFB1, p < 0.05). Importantly, the expression of these genes was partially or even totally suppressed by the pre-exposure of cells to HT (GAPDH, p < 0.01 for HT + CoCl2 vs. CoCl2; expression of HIF1A, IL6 and TGFB1 for HT + CoCl2 was similar to that of control). Conclusion Our data supports the potential for a multiplicity of preventive effects of HT, providing antioxidant, anti-inflammatory and anti-fibrotic defenses to renal cells under hypoxic conditions. Importantly, the development of safe and effective therapeutic approaches based on phytochemicals such as HT, may present substantial advantages for renal patients over synthetic drugs, including fewer side effects, significantly lower price, and ease of administration in the form of dietary supplements. Acknowledgments This work was supported by Applied Molecular Biosciences Unit (UCIBIO), financed by national funds from FCT/MCTES (UIDB/04378/2020), by North Portugal Regional Coordination and Development Commission (CCDR-N)/NORTE2020/Portugal 2020 (Norte-01-0145-FEDER-000024), and co-financed by FCT/MCTES (PTDC/OCE-ETA/32492/2017) and FEDER/COMPETE 2020 (POCI-01-0145-FEDER-032492).

Calcium signaling in cultured human and rat duodenal enterocytes

1998 ◽  
Vol 275 (2) ◽  
pp. G296-G304 ◽  
Author(s):  
Catherine S. Chew ◽  
Bengt Säfsten ◽  
Gunnar Flemström
Keyword(s):  
Extracellular Matrix ◽  
Calcium Signaling ◽  
Primary Culture ◽  
Calcium Concentration ◽  
Duodenal Mucosa ◽  
Transient Increase ◽  
Active Movement ◽  
Culture Model ◽  
Columnar Cells ◽  
Sustained Increase

Vagal stimuli increase duodenal mucosal[Formula: see text] secretion and may provide anticipatory protection against acid injury, but duodenal enterocyte (duodenocyte) responses and cholinoceptor selectivity have not been defined. We therefore developed a stable primary culture model of duodenocytes from rats and humans. Brief digestion of scraped rat duodenal mucosa or human biopsies with collagenase/dispase yielded cells that attached to the extracellular matrix Matrigel within a few hours of plating. Columnar cells with villus enterocyte morphology that exhibited spontaneous active movement were evident between 1 and 3 days of culture. Rat duodenocytes loaded with fura 2 responded to carbachol with a transient increase in intracellular calcium concentration ([Ca2+]i), with an apparent EC50 of ∼3 μM. In a first type of signaling pattern, [Ca2+]ireturned to basal or near basal values within 3–5 min. In a second type, observed in cells with enlarged vacuoles characteristic of crypt cell morphology, the initial transient increase was followed by rhythmic oscillations. Human duodenocytes responded with a more sustained increase in [Ca2+]i, and oscillations were not observed. Rat as well as human duodenocytes also responded to CCK-octapeptide but not to vasoactive intestinal polypeptide. Equimolar concentrations (100 nM) of the subtype-independent muscarinic antagonist atropine and the M3 antagonist 4-diphenylacetoxy- N-methylpiperidine methiodide prevented the response to 10 μM carbachol, whereas the M1 antagonist pirenzepine and the M2 antagonists methoctramine and AF-DX 116BS had no effect at similar concentrations. Responses in rat and human duodenocytes were similar. A new agonist-sensitive primary culture model for rat and human duodenocytes has thus been established and the presence of enterocyte CCK and muscarinic M3 receptors demonstrated.

3D Primary Culture Model to Study Human Mammary Development

2017 ◽  
pp. 139-147 ◽  
Author(s):  
Daniel H. Miller ◽  
Ethan S. Sokol ◽  
Piyush B. Gupta
Keyword(s):  
Primary Culture ◽  
Mammary Development ◽  
Culture Model

Effects of Caffeine on Intervertebral Disc Cell Viability in a Whole Organ Culture Model

2017 ◽  
Vol 17 (10) ◽  
pp. S125-S126
Author(s):  
Benjamin T. Raines ◽  
James T. Stannard ◽  
Olivia E. Stricklin ◽  
Aaron M. Stoker ◽  
Theodore J. Choma ◽  
...  
Keyword(s):  
Intervertebral Disc ◽  
Organ Culture ◽  
Cell Viability ◽  
Disc Cell ◽  
Culture Model

scholarly journals Osteoblast differentiation of human bone marrow cells under continuous and discontinuous treatment with dexamethasone

2005 ◽  
Vol 16 (2) ◽  
pp. 156-161 ◽  
Author(s):  
Márcio Mateus Beloti ◽  
Adalberto Luiz Rosa
Keyword(s):  
Bone Marrow ◽  
Cell Proliferation ◽  
Cell Viability ◽  
Protein Content ◽  
Primary Culture ◽  
Total Protein ◽  
Osteoblast Differentiation ◽  
Total Protein Content ◽  
First Passage ◽  
Alp Activity

Dexamethasone (Dex) has been shown to induce osteoblast differentiation in several cell culture systems. This study investigated the effect of continuous and discontinuous treatment with Dex on osteoblast differentiation of human bone marrow stromal cells (BMSC). Primary culture and first passage were cultured in media with or without Dex 10-7 M. During the culture period, cells were incubated at 37ºC in humidified atmosphere of 5% CO2 and 95% air. At 7, 14, and 21 days, cell proliferation, cell viability, total protein content, alkaline phosphatase (ALP) activity and bone-like formation were evaluated. Data were compared by two-way analysis of variance. Dex did not affect cell viability and total protein content, but reduced cell number. ALP activity and bone-like formation increased when only first passage or both primary culture and first passage were treated with Dex, in comparison to the groups that did not have contact with Dex after first passage. The results of this study indicate that, for human BMSC, continuous presence of Dex did not appear to be required for development of the osteoblast phenotype, but Dex must be present after first passage to allow osteoblast differentiation expressed by reduced cell proliferation and increased ALP activity and bone-like formation.

Microfluidic primary culture model of the lower motor neuron–neuromuscular junction circuit

2013 ◽  
Vol 218 (2) ◽  
pp. 164-169 ◽  
Author(s):  
Katherine A. Southam ◽  
Anna E. King ◽  
Catherine A. Blizzard ◽  
Graeme H. McCormack ◽  
Tracey C. Dickson
Keyword(s):  
Neuromuscular Junction ◽  
Motor Neuron ◽  
Primary Culture ◽  
Lower Motor Neuron ◽  
Culture Model

scholarly journals Characterization of folate uptake by choroid plexus epithelial cells in a rat primary culture model

2007 ◽  
Vol 104 (6) ◽  
pp. 1494-1503 ◽  
Author(s):  
Jan B. Wollack ◽  
Benedette Makori ◽  
Stuti Ahlawat ◽  
Rajeth Koneru ◽  
Sonia C. Picinich ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Primary Culture ◽  
Choroid Plexus ◽  
Culture Model ◽  
Choroid Plexus Epithelial

scholarly journals Primary culture model of peroxisome proliferator-activated receptor ? activity in prostate cancer cells

2003 ◽  
Vol 196 (1) ◽  
pp. 131-143 ◽  
Author(s):  
Yue Xu ◽  
Sunita Iyengar ◽  
Richard L. Roberts ◽  
Scott B. Shappell ◽  
Donna M. Peehl
Keyword(s):  
Prostate Cancer ◽  
Cancer Cells ◽  
Primary Culture ◽  
Receptor Activity ◽  
Peroxisome Proliferator ◽  
Prostate Cancer Cells ◽  
Peroxisome Proliferator Activated Receptor ◽  
Culture Model
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