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Nahco3

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NaHCO3 is a chemical compound that serves as a buffer solution. It is a white, crystalline solid that dissolves in water to produce a basic, alkaline solution. NaHCO3 is commonly used in various laboratory applications to maintain pH levels and control chemical reactions.

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Lab products found in correlation

D glucose, by Harvard Bioscience (9 mentions) Fetal bovine serum (fbs), by Harvard Bioscience (5 mentions) Stable glutamine, by Harvard Bioscience (5 mentions) Na pyruvate, by Harvard Bioscience (3 mentions) Na pyruvat, by Harvard Bioscience (2 mentions) Penicillin streptomycin, by Harvard Bioscience (2 mentions) Penicillin, by Merck Group (2 mentions) A549 cell, by American Type Culture Collection (2 mentions) Qubit 3.0 fluorometer, by Thermo Fisher Scientific (2 mentions) L glutamine, by Merck Group (2 mentions) Nuclisens easymag, by bioMérieux (2 mentions) Qubit dsdna high sensitivity assay kit, by Thermo Fisher Scientific (2 mentions) Penicillin, by Harvard Bioscience (2 mentions) Streptomycin, by Harvard Bioscience (2 mentions) L glutamine, by Harvard Bioscience (2 mentions) Hepes, by Harvard Bioscience (2 mentions) Non essential amino acids (neaa), by PAN Biotech (1 mentions) L glutamine, by PAN Biotech (1 mentions) Penicillin streptomycin, by Merck Group (1 mentions) Trypsin edta, by Harvard Bioscience (1 mentions)

19 protocols using nahco3

1

Immortalized 16HBE14o- Epithelial Cell Line

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The immortalized
16HBE14o- epithelial cell line is derived from transformed bronchial
epithelial cells of a 1-year-old heart–lung transplant patient.29 (link) Cultivation of the cells was carried out in
eukaryotic minimal essential medium (eMEM): 1× MEM with Earle’s
salts with 2.2 g/L NaHCO3 (Biochrom AG, Berlin, Germany)
supplemented with 10% (v/v) fetal calf serum (FCS; Biochrom AG), 2%
(v/v) l-glutamine (200 mM, PAN-Biotech GmbH), and 1% (v/v)
nonessential amino acids (100×, PAN-Biotech GmbH). The cells
were cultured in 10 cm plates at 37 °C and 5% CO2 in
a humid atmosphere and were kept for no more than 15 passages.
The seeding of the cells was done at a density of 1 × 105 cells/cm2 over a 12 mm Transwell polyester membrane
with 0.4 μm pore size (Corning, Schnelldorf, Germany) to promote
the polarization of the cell layer. Cells were cultured for 3 or 11
days, depending on the desired condition for infection. The volume
of medium on the apical side was 400 μL, and it was 1300 μL
on the basal side. The medium was exchanged every second day until
day eight after which the exchange was done daily.
Palma Medina L.M., Becker A.K., Michalik S., Surmann K., Hildebrandt P., Gesell Salazar M., Mekonnen S.A., Kaderali L., Völker U, & van Dijl J.M. (2020). Interaction of Staphylococcus aureus and Host Cells upon Infection of Bronchial Epithelium during Different Stages of Regeneration. ACS Infectious Diseases, 6(8), 2279-2290.
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2

Glioblastoma Cell Culture Protocol

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Two human glioblastoma cell lines were used. U-251 MG human brain glioblastoma cell line was obtained from CLS (Heidelberg, Germany) and U-373 MG was a generous gift from Dr. Bardenheuer (Essen, Germany). The cell lines U-251 and U-373 were routinely grown in Dulbecco’s modified eagle medium (DMEM) with 4.5 g/l d-Glucose, 3.7 g/l NaHCO3, stable glutamine and Na-pyruvat (Biochrom AG, FG 0445, Germany). Media were supplemented with 10% sterile fetal bovine serum (Biochrom AG, S 0115) and 1% penicillin–streptomycin (Sigma-Aldrich, P0781). Cell lines were maintained at 37°C, 5% CO2, and 90% humidity.
Krcek R., Matschke V., Theis V., Adamietz I.A., Bühler H, & Theiss C. (2017). Vascular Endothelial Growth Factor, Irradiation, and Axitinib Have Diverse Effects on Motility and Proliferation of Glioblastoma Multiforme Cells. Frontiers in Oncology, 7, 182.
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3

Microcontact Printing with Engineered NIH 3T3 Cells

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We used NIH 3T3 (ATCC CRL1658) fibroblasts for experiments involving microcontact printing. The NIH 3T3 cell culture was a gift from Louis Lim (Institute of Molecular and Cell Biology, ASTAR Singapore). For long-term experiments we employed a genetically modified variant termed NIH 3T3 X2 [39 (link)]. Compared with standard 3T3 cells, an enlarged fraction of X2 cells exhibited CDR formation. Cells were grown under standard conditions in Dulbecco’s MEM containing 3.7 g/L NaHCO3, 4.5 g/L D-Glucose (Biochrom), 100 µg/ml Penicillin/Streptomycin (PAA), and 10% Fetal Bovine Serum (Biochrom). Cells were split at 80% confluency using Trypsin/EDTA (Biochrom). Cells were mycoplasma free.
Bernitt E., Koh C.G., Gov N, & Döbereiner H.G. (2015). Dynamics of Actin Waves on Patterned Substrates: A Quantitative Analysis of Circular Dorsal Ruffles. PLoS ONE, 10(1), e0115857.
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4

Cultivation and Treatment of Glioblastoma Cells

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U-87 MG was obtained from the American Type Culture Collection (ATCC) (Manassas, USA) and U-251 MG from CLS (Heidelberg, Germany). Cells were cultivated in Dulbecco's modified eagle medium (DMEM) containing 4.5 g/L D-Glucose, 3.7 g/L NaHCO3, stable glutamine and Na-Pyruvat (Biochrom AG, FG 0445, Germany) and supplemented with 10% sterile fetal bovine serum (Biochrom AG, S 0115) and 1% penicillin (Sigma-Aldrich). Cells were maintained at 37°C, 5% CO2 and 90% air moisture. Subcultures were made on 12 mm Ø cover slips to apply VEGF, axitinib or radiation on the confluent cell monolayer.
Krcek R., Latzer P., Adamietz I.A., Bühler H, & Theiss C. (2017). Influence of vascular endothelial growth factor and radiation on gap junctional intercellular communication in glioblastoma multiforme cell lines. Neural Regeneration Research, 12(11), 1816-1822.
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5

Hormone Receptor-Positive Breast Cancer Cell Culture

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MCF-7 cells were chosen for their hormone receptor-positive attributes, reflecting the luminal breast cancer subtype. Their estrogen and progesterone receptor presence aligns with our hormone-related focus. Our selection was guided by their relevance to our objectives, well-documented status, and specific interest in MTE's impact on this hormone receptor-positive subtype. Additionally, our choice of MCF-7 cells was influenced by the existing data on green tea phenol's interaction with ERα, while information about ERβ interactions remains limited.
MCF-7 cells were cultured on an 80% monolayer in a cell culture bottle. Dulbecco's modified Eagle medium (DMEM; 3.7 g/l NaHCO3, 4.5 g/l d-glucose, 1.028 g/l stable glutamine, and sodium pyruvate; Biochrom, Berlin, Germany) supplemented with 10% heat-inactivated fetal calf serum (FCS; Biochrom, Berlin, Germany) was used for cultivation. The cells were incubated with atmospheric concentrations of CO2 of 5% at 37 °C and were then trypsinized and counted for further use.
Keckstein S., Tilgener C., Jeschke U., Hofmann S., Vilsmaier T., Keilmann L., Heidegger H., Kaltofen T., Batz F., Mahner S, & Schröder L. (2023). Effects of matcha tea extract on cell viability and estrogen receptor-β expression on MCF-7 breast cancer cells. Archives of Gynecology and Obstetrics, 309(4), 1509-1514.
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6

Trophoblast Cell Line Stimulation Assay

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The choriocarcinoma cell lines BeWo (ECACC, Salisbury, UK) and JEG-3 (ATCC), which are useful models of human trophoblasts, were used for the study. The cells were cultured in DMEM (3.7 g/L NaHCO3, 4.5 g/L d-glucose, 1.028 g/L stable glutamine, and Na-Pyruvate; Biochrom, Berlin, Germany). 10% heat-inactivated FCS was added to the medium, and the solution was incubated at an atmospheric CO2 concentration of 5% and at 37°C.
BeWo and JEG-3 cells were separately grown on sterile 12 multiwell slides at a density of 500,000 cells/mL DMEM with 10% foetal cow serum. After 4 h, medium was changed to pure DMEM. The cells were stimulated with 0.01 nM or 0.1 nM triiodothyronine (T3) (Sigma T2877; Lot:106K1157V, Sigma-Aldrich), thyronamine (T0AM) (Fluka 80345, Lot: BCBL2185V, Buchs, Switzerland), 3-Iodothyronamine (T1AM) (Cayman Chemical) and RO5203548 (Glixx Laboratories, Hopkinton, MA, USA) for 2 h (PCR samples) and 48 h (Immunocytochemistry and Western blot samples), respectively. Control cells were incubated without stimulants. As a control solvent, the following buffers were used: The stimulant T3 has been diluted in 1 M NaOH in DMEM (Dulbecco’s Modified Eagle Medium) and T1AM and RO5203548 were diluted in DMSO (Dimethylsulfoxide).
Stavrou S., Gratz M., Tremmel E., Kuhn C., Hofmann S., Heidegger H., Peryanova M., Hermelink K., Hutter S., Toth B., Mayr D., Mahner S., Jeschke U, & Vattai A. (2018). TAAR1 induces a disturbed GSK3β phosphorylation in recurrent miscarriages through the ODC. Endocrine Connections, 7(2), 372-384.
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7

Cell Culture and DNA Extraction Protocol

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Cell cultures were performed in A549 cells purchased from the American Type Culture Collection (Manassas, VA, USA) and were maintained in Earle’s minimal essential medium (MEM) (Biochrom GmbH, Germany) supplemented with 2.2 g/l NaHCO3 (Biochrom), 1% l-glutamine (Merck, Germany), 1% penicillin (10,000 U/ml) and streptomycin (10,000 μg/ml) (Biochrom), 1% fungizol (CPS Cito Pharma Services, Switzerland), and 1% heat inactivated Fetal Bovine Serum (FBS) (Biochrom) at 37 °C and 5% CO2. The samples inoculated in A549 cells were incubated at 37 °C for 3–7 days or until a cytopathic effect was observed. After cytopathic effect was confirmed, the positive cell culture supernatant was subjected to DNA extraction. DNA viral extraction from 200 µl supernatant of the cell cultured HAdV positive samples was automatically performed using the NUCLISENS easyMAG (bioMérieux, Geneva, Switzerland) extractor, as per manufacturer’s instructions. After DNA extraction, the DNA concentration of each sample was measured using the Qubit dsDNA high sensitivity assay kit on the Qubit 3.0 Fluorometer (ThermoFisher Scientific, Zug, Switzerland) as per manufacturer's protocol prior to whole genome amplification (WGA).
Akello J.O., Kamgang R., Barbani M.T., Suter-Riniker F., Aebi C., Beuret C., Paris D.H., Leib S.L, & Ramette A. (2021). Genomic analyses of human adenoviruses unravel novel recombinant genotypes associated with severe infections in pediatric patients. Scientific Reports, 11, 24038.
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8

Culturing SKOV3 ovarian cancer cells

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The human ovarian cancer cell line SKOV3 was purchased from the European Collection of Authenticated Cell Cultures (ECACC). SKOV3 was cultured in McCoy’s 5A medium (modified), w: L-glutamine, w: 2.2 g/L NaHCO3 (Biochrom), supplemented with 15% FBS-fetal bovine serum (Gibco) and 1% penicillin–streptomycin (P/S), as recommended by the suppliers.
Constantinescu D.R., Sorop A., Ghionescu A.V., Lixandru D., Herlea V., Bacalbasa N, & Dima S.O. (2024). EM-transcriptomic signature predicts drug response in advanced stages of high-grade serous ovarian carcinoma based on ascites-derived primary cultures. Frontiers in Pharmacology, 15, 1363142.
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9

Thyroid Hormone Signaling in Breast Cancer

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Ductal breast cancer cell lines MCF7 and T47D (both ECACC, Salisbury, UK) were used for the experiments. The cells were cultured in DMEM (3.7 g/L NaHCO3, 4.5 g/L d-glucose, 1.028 g/L stable glutamine, and Na-pyruvate; Biochrom, Berlin, Germany) adding 10% heat-inactivated fetal calf serum (FCS; Biochrom) to the medium, and the solution was incubated at an atmospheric CO2 concentration of 5% and at a temperature of 37°C.
For the PCR and Western blot stimulation, MCF7 and T47D cells were separately grown on sterile 12 multiwell slides at a density of 500,000 cells/mL DMEM with 10% FCS. Medium was changed after 4 hours to pure DMEM without FCS. After 16 hours, each group of cells was stimulated with 0.01 or 0.1 nM T1AM (Cayman Chemical, Ann Arbor, USA), T3 (Sigma-Aldrich Co., St Louis, MO, USA), and tetraiodothyroacetic acid (Tetrac) (Sigma-Aldrich Co.) for 2 hours in case of TaqMan® PCR experiments. For Western blot lysates, each group of cells was stimulated with 1 or 10 nM T1AM, T3, and Tetrac for 24 hours. 10 µg/mL of estradiol (Sigma-Aldrich Co.) was added to T47D cells, which were stimulated with 10 nM T1AM.
Control cells were incubated under the same conditions without stimulants.
Tremmel E., Hofmann S., Kuhn C., Heidegger H., Heublein S., Hermelink K., Wuerstlein R., Harbeck N., Mayr D., Mahner S., Ditsch N., Jeschke U, & Vattai A. (2019). Thyronamine regulation of TAAR1 expression in breast cancer cells and investigation of its influence on viability and migration. Breast Cancer : Targets and Therapy, 11, 87-97.
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10

Breast Cancer Cell Line Stimulation Assay

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The cell lines MCF7 and T47D were used as models for the ductal breast carcinoma. The cells were cultured in DMEM (3.7 g/L NaHCO3, 4.5 g/L D-glucose, 1.028 g/L stable glutamine, and Na-Pyruvate; Biochrom, Berlin, Germany). After adding 10% heat-inactivated fetal calf serum (FCS; Biochrom) to the medium the solution was incubated at an atmospheric CO2 concentration of 5% and at 37 °C in an incubator to lyse.
MCF7 and T47D cells were separately grown in sterile 12 multi-well slides at a density of 500,000 cells/mL DMEM with 10% FCS. The medium was changed after 4 h to pure DMEM without any FCS. After 20 h, the cells were stimulated with 1 μM epinephrine (Sigma-Aldrich Chemie GmbH, Munich, Germany). The cells were stimulated for 6 h with 10 μM epinephrine in the case of Western blot samples. Control cells were incubated in parallel without any stimulants.
Tremmel E., Kuhn C., Kaltofen T., Vilsmaier T., Mayr D., Mahner S., Ditsch N., Jeschke U, & Vattai A. (2020). L-Dopa-Decarboxylase (DDC) Is a Positive Prognosticator for Breast Cancer Patients and Epinephrine Regulates Breast Cancer Cell (MCF7 and T47D) Growth In Vitro According to Their Different Expression of Gi- Protein- Coupled Receptors. International Journal of Molecular Sciences, 21(24), 9565.
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