Glycol chitosan (GC, degree of polymerization ≥400, #G7753), cerium (III) chloride, RPMI medium 1640 (1×), DMEM/F12+GlutaMAX™-I (1×) medium, Improved MEM (Richter’s modified) medium, antibiotic-antimycotic solution (100×), and 0.5% Trypsin-EDTA (1×) were purchased from Gibco Life Technologies (Grand Island, NY, USA). Fetal bovine serum (FBS) was purchased from Invitrogen (Carlsbad, CA, USA). All experiments were carried out and animals were maintained in accordance with the Association for Research in Vision and Ophthalmology (ARVO) statement for the use of animals in ophthalmic and vision research and the guidelines of the University of North Carolina at Chapel Hill Animal Care and Use Committee.
Antibiotic antimycotic solution 100
Manufactured by Thermo Fisher Scientific
Sourced in United States
Antibiotic/antimycotic solution (100×) is a concentrated mixture of antibiotics and antifungal agents used to prevent microbial contamination in cell culture and other laboratory applications. It contains a combination of penicillin, streptomycin, and amphotericin B at 100 times the typical working concentration.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (15 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (4 mentions)
Dulbecco s modified eagle s medium dmem, by Thermo Fisher Scientific (3 mentions)
Methyl thiazolyl tetrazolium (mtt), by HiMedia (2 mentions)
Chloramphenicol, by Merck Group (2 mentions)
α mem, by Thermo Fisher Scientific (2 mentions)
Lipofectamine rnaimax transfection reagent, by Thermo Fisher Scientific (2 mentions)
Penicillin, by Thermo Fisher Scientific (2 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions)
Collagenase type 4, by Merck Group (1 mentions)
Dulbecco s modified eagle s medium dmem f12, by Thermo Fisher Scientific (1 mentions)
Doxorubicin hydrochloride solution, by Merck Group (1 mentions)
Phosphate buffered saline (pbs), by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by HiMedia (1 mentions)
Tetracyclin, by Merck Group (1 mentions)
Rifamycin, by Merck Group (1 mentions)
Flutamide, by Merck Group (1 mentions)
Dnase 1, by New England Biolabs (1 mentions)
Escort 4 reagent, by Merck Group (1 mentions)
Hydrocortisone, by HiMedia (1 mentions)
20 protocols using antibiotic antimycotic solution 100
1
Glycol Chitosan-based Cell Culture
2
Isolation and Characterization of Endometrial Stromal Cells
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The primary cultures of the non-endometriosis and endometriosis endometrial stromal cells and endometriotic lesion stromal cells were performed as described previously [75 (link)]. Briefly, the tissues were digested for 40 min at 37 °C with 0.5% collagenase type IV (Sigma–Aldrich, Saint Louis, MO, USA). The cells were isolated in Dulbecco’s modified Eagle’s medium (DMEM, Gibco Laboratories, Grand Island, NY, USA). A 250-mm sieve (Sigma–Aldrich, Saint Louis, MO, USA) was used to filter the cell suspensions. A second sieve with 35-mm mesh was used to retain the clustered cells, such as epithelial cells. The flow-through with the single cells was seeded in flasks. Culture and expansion were performed in DMEM supplemented with 15% fetal bovine serum and 1% antibiotic/antimycotic solution (100×) (Gibco Laboratories, Grand Island, NY, USA) at 37 °C in a 5% CO2 incubator. After six passages, the absence of epithelial cell contamination was assessed by immunostaining the cells with anti-vimentin as a specific marker of stromal cells and anti-E-cadherin, a specific marker of epithelial cells.
3
Cell Viability Assay with Withaferin A
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PBS (pH = 7.2, 1x), 0.25% trypsin–EDTA (1x), Dulbecco's Modified Eagle's Medium DMEM/F-12 (1x), 0.4% trypan blue, and antibiotic/antimycotic solution (100×) were obtained from Gibco, Life Technologies; whereas fetal bovine serum (FBS) and MTT were obtained from Himedia. Doxorubicin hydrochloride solution was obtained from Sigma Chemical Co. (St. Louis, MO, USA) and dimethyl sulfoxide (DMSO) from Calbiochem. Withaferin A standard was from Natural Remedies Ltd. Veerasandra, Bangalore-100. All HPLC grade reagents were used in HPLC. All other chemicals were of analytical grade.
4
Fetal Bovine Serum and Antibiotic Optimization
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Fetal bovine serum (FBS) and an antibiotic-antimycotic
solution (100×) were purchased from the GIBCO (GIBCO, BRL, Inchinnman
UK). Escort IV reagent, Dithiothreitol (DTT), anti-Rabbit-IgG-cy-3,
Hoechst 33258, TRI-Reagent, testosterone, dihydrotestosterone (DHT),
β-estradiol, and flutamide were purchased from Sigma-Aldrich
(St. Louis, MO, USA). Hydrocortisone, MTT (3-(4,5-dimethyl-2-thiazolyl)
2,5diphenyl-2H-tetrazolium bromide), and cell culture media were purchased
from Himedia (Mumbai India). DNaseI was purchased from NEB, England.
Oxytetracyclin (catalog number-75966), chloramphenicol (catalog number-C0378),
oxacillin (catalog number-28221), streptomycin (catalog number-S6501),
tetracyclin (catalog number-T3258), rifamycin (catalog number-R3501),
ciprofloxacin (catalog number-17850), rotenone (catalog number-R8875),
and BPA (catalog number-239658) purchased from Sigma-Aldrich (St.
Louis, MO, USA) were a kind gift from Prof. S. P. Singh, Banaras Hindu
University, Varanasi, India.
solution (100×) were purchased from the GIBCO (GIBCO, BRL, Inchinnman
UK). Escort IV reagent, Dithiothreitol (DTT), anti-Rabbit-IgG-cy-3,
Hoechst 33258, TRI-Reagent, testosterone, dihydrotestosterone (DHT),
β-estradiol, and flutamide were purchased from Sigma-Aldrich
(St. Louis, MO, USA). Hydrocortisone, MTT (3-(4,5-dimethyl-2-thiazolyl)
2,5diphenyl-2H-tetrazolium bromide), and cell culture media were purchased
from Himedia (Mumbai India). DNaseI was purchased from NEB, England.
Oxytetracyclin (catalog number-75966), chloramphenicol (catalog number-C0378),
oxacillin (catalog number-28221), streptomycin (catalog number-S6501),
tetracyclin (catalog number-T3258), rifamycin (catalog number-R3501),
ciprofloxacin (catalog number-17850), rotenone (catalog number-R8875),
and BPA (catalog number-239658) purchased from Sigma-Aldrich (St.
Louis, MO, USA) were a kind gift from Prof. S. P. Singh, Banaras Hindu
University, Varanasi, India.
5
Cytotoxicity of Cancer Cells Exposed to ME
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The human pancreatic cancer cell line BxPC-3 (ATCC CRL-1687) was purchased from the American Type Culture Collection (USA). Roswell Park Memorial Institute (RPMI) medium, fetal bovine serum (FBS), antibiotic–antimycotic solution (100×), 0.25% trypsin-ethylene diamine tetraacetic acid solution, and phosphate-buffered saline (PBS) were procured from Gibco (USA). The cells were cultured in RPMI medium containing 10%(v/v) FBS and 1% (w/v) antibiotic–antimycotic at 37°C in a 5% (v/v) CO2 incubator. Human mesenchymal stem cells (hMSCs) were purchased from Lonza (USA) and cultured in StemMACSTM MSC expansion media (Miltenyi Biotec, Germany). Cells (5 × 103 cells/well) were seeded in 96-well plates in a medium containing 5%FBS and 1% antibiotics. After 24h, the medium was replaced with fresh medium containing each ME concentration for 24, 48, and 72 h. At each time point, the number of viable cells was counted under a microscope (Nikon Eclipse TE300; Nikon, Japan).
6
Culture of OSCA-8 Cells
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OSCA-8 cells (Kerafast, Inc., Boston, MA, USA) were maintained in Dulbecco’s Modified Eagle’s Medium (Santa Cruz Biotechnology Inc., USA) supplemented with 10% fetal calf serum and antibiotic-antimycotic solution (100×, Gibco, Thermo Fisher Scientific Inc., USA). In addition, cells were maintained at 37°C in a humidified 5% CO2 atmosphere.
7
Mesenchymal Stem Cell Culture and Differentiation
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Dulbecco’s modified Eagle medium (DMEM) (1.0 g/L glucose), Dulbecco’s phosphate-buffered saline (DPBS), and trypsin-ethylenediaminetetraacetic acid (EDTA) solution were purchased from Nacalai Tesque (Kyoto, Japan). Mesenchymal Stem Cell Growth Medium Bullet Kit™ (PT-3001), hMSC Osteogenic Differentiation Medium Bullet Kit™ (PT-3002), and hMSC Adipogenic Differentiation Medium Bullet Kit™ (PT-3004) were purchased from Lonza (Walkersville, MD, USA). Fetal bovine serum (FBS) and antibiotic-antimycotic solution 100× were purchased from Gibco (Waltham, MA, USA).
8
Culturing Human Fibroblasts and Mesenchymal Stem Cells
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Human embryo fibroblasts (HFs) (OUMS-36T-1) were purchased from JCRB Cell Bank (Osaka, Japan) and cultured in a 37 °C and 5% CO2 incubator with DMEM (Nacalai Tesque) supplemented with 10% FBS (Gibco) and 5% antibiotic-antimycotic solution 100× (Gibco). The medium was changed every two days. HFs were passaged when they became sub-confluent (approximately 70% confluent).
Normal bone marrow-derived human mesenchymal stem cells (hMSCs) were purchased from Lonza and cultured in a 37 °C and 5% CO2 incubator with hMSC basal medium containing SingleQuots™ Supplements and Growth Factors (PT-3001, Lonza). The medium was changed every two days. HFs were passaged when were sub-confluent (approximately 80% confluent). hMSCs (passage 3–5) were used in differentiation experiments.
Normal bone marrow-derived human mesenchymal stem cells (hMSCs) were purchased from Lonza and cultured in a 37 °C and 5% CO2 incubator with hMSC basal medium containing SingleQuots™ Supplements and Growth Factors (PT-3001, Lonza). The medium was changed every two days. HFs were passaged when were sub-confluent (approximately 80% confluent). hMSCs (passage 3–5) were used in differentiation experiments.
9
Anatase TiO₂ NPs Cytotoxicity Assay
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Dulbecco’s modified Eagle’s medium (DMEM) high glucose, 0.25% trypsin-EDTA solution, Antibiotic Antimycotic Solution (100×), and fetal bovine serum (FBS) were acquired from Gibco BRL (Grand Island, NY, USA). Cell culture consumables were purchased from Corning (Corning, NY, USA). Flow cytometry reagents were provided by Becton-Dickinson Immunocytometry Systems (San Jose, CA, USA). H2DCFDA was purchased from Molecular Probes, Invitrogen (Carlsbad, CA, USA). CytoTox 96 Non-radioactive cytotoxicity assay was from Promega (Madison, WI, USA). Western blot reagents were from Bio-Rad (Hercules, CA, USA). Autophagy detection kit was purchased from abcam (Cambridge, MA, USA). Anatase TiO2 NPs 25 nm and other chemicals were obtained from Sigma-Aldrich (St. Louis, MO, USA).
10
Propagation and Cultivation of PEDV Strains
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Vero cells (ATCC CCL-81) were cultured in alpha-minimum essential medium (α-MEM; Invitrogen, Carlsbad, CA) with 5% fetal bovine serum (FBS; Invitrogen) and antibiotic-antimycotic solution (100×; Invitrogen) and maintained at 37 °C in a humidified 5% CO2 incubator. The highly virulent Korean PEDV G2b strain KOR/KNU-141112/2014 was isolated and propagated in Vero cells as described previously (Lee et al., 2015 (link)). Viral stock from fifth passage cell culture (KNU-141112-P5) was prepared and used as the challenge virus in this study. The fully attenuated S DEL5/ORF3 strain was cultivated in Vero cells as described previously (Lee et al., 2017 (link)) and prepared as the live vaccine stock (T317PEDL01; 105.0 TCID50/ml) according to instructions from ChoongAng Vaccine Laboratories (CAVAC; Daejeon, South Korea).
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