ARPE-19 cells (CRL-2302, ATCC) were cultured in DMEM/F12 (Corning) with 10% FBS (Omega Scientific) and 1% PSQ (ThermoFisher). Cells were kept at 37°C with 5% CO2, and passages were performed every 3-4 days, or when confluent. For experiments, ARPE-19 cells were plated at confluence (unless otherwise noted), typically at a concentration of ~100,000 cells/cm2, which equates to ~200,000 cells/mL. HEK-293T cells (Life Technologies) were cultured in DMEM High Glucose (4.5 g/L) supplemented with 10% FBS and 1% PSQ. Mouse NIH-3T3 fibroblasts (CRL-1658, ATCC) were maintained in DMEM high-glucose with 10% calf serum and 1% PSQ. Primary dermal fibroblasts (PCS-201-012, ATCC) were maintained in either low glucose DMEM with 10% FBS and 1% PSQ, or fibroblast growth media (Fibroblast Basal Medium [PCS-201-012, ATCC] supplemented with Low-Serum Fibroblast Growth Kit [PCS-201-041, ATCC]. Cells were periodically confirmed to be free of mycoplasma contamination using the Universal Mycoplasma Detection Kit (30-1012k, ATCC).
Pcs 201 012
Manufactured by American Type Culture Collection
Sourced in United States
The PCS-201-012 is a laboratory equipment product from American Type Culture Collection. It is designed for cell culture applications, but a detailed description of its core function cannot be provided while maintaining an unbiased and factual approach. More information would be required to describe the product accurately without interpretation or extrapolation.
Automatically generated - may contain errors
Lab products found in correlation
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (7 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (6 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (5 mentions)
High glucose dmem, by Thermo Fisher Scientific (3 mentions)
Fibroblast growth kit low serum, by American Type Culture Collection (2 mentions)
Dulbecco modified eagle medium (dmem), by American Type Culture Collection (2 mentions)
Pcs 201 030, by American Type Culture Collection (2 mentions)
Pcs 201 041, by American Type Culture Collection (2 mentions)
Hb 8065, by American Type Culture Collection (2 mentions)
Ccl 81, by American Type Culture Collection (2 mentions)
Pcs 201 010, by American Type Culture Collection (2 mentions)
L glutamine, by Thermo Fisher Scientific (2 mentions)
Fetal calf serum (fcs), by Thermo Fisher Scientific (2 mentions)
Pcs 200 013, by American Type Culture Collection (2 mentions)
Wm1552c, by Rockland Immunochemicals (2 mentions)
Pcs 200 010, by American Type Culture Collection (2 mentions)
Crl 2522, by American Type Culture Collection (2 mentions)
Htb 26, by American Type Culture Collection (2 mentions)
Low serum growth supplement, by Thermo Fisher Scientific (2 mentions)
Mda mb 231, by American Type Culture Collection (2 mentions)
63 protocols using pcs 201 012
1
Cell Culture Protocols for ARPE-19, HEK-293T, NIH-3T3, and Primary Fibroblasts
2
Cell Line Cultivation Protocol for Cancer Research
3
Synthesis of Gold Nanoparticles
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Tetrachloroauric(III) acid
trihydrate (HAuCl4·3H2O), citric acid (CA),
sodium tricitrate dihydrate, and ethylene diamine (EDA) were obtained
from Sigma-Aldrich and used without further purification. Prior to
experiments, all glassware was cleaned with aqua regia and washed
with distilled water. Adenocarcinoma human alveolar basal epithelial
cells (A549, CCL-185, ATCC) and HDF cells (HDFa, PCS-201-012, ATCC)
cell lines were purchased from the American Type Culture Collection.
trihydrate (HAuCl4·3H2O), citric acid (CA),
sodium tricitrate dihydrate, and ethylene diamine (EDA) were obtained
from Sigma-Aldrich and used without further purification. Prior to
experiments, all glassware was cleaned with aqua regia and washed
with distilled water. Adenocarcinoma human alveolar basal epithelial
cells (A549, CCL-185, ATCC) and HDF cells (HDFa, PCS-201-012, ATCC)
cell lines were purchased from the American Type Culture Collection.
4
Fibroblast Viability Assay with Conditioned Media
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The human primary dermal fibroblast cell lines (catalog #PCS-201-012, ATCC, VA, USA) were purchased and then cultured and expanded in standard media supplemented with 10% FBS and 1% penicillin–streptomycin and used for the experiments in passages 3–5. When the fibroblasts reached 70% confluence, they were trypsinized and subcultured to culture flasks.
The viability of the HDFs after treatment with gel-CM was assessed using the Cell Counting Kit-8 (CCK-8) assay (Dojindo, Japan). The cells (1 × 104/well) were plated on 96-well plates for 24 h, followed by treatment with the gel-CM from the serum-free media (gel-CM), serum-deprivation media (2% FBS), and standard media (10% FBS). After treatments for 24, 48, and 72 h, the cell viability was measured using a CCK-8 assay kit (Dojindo, Japan) according to the manufacturer’s instructions, respectively. All experiments were performed three times.
The viability of the HDFs after treatment with gel-CM was assessed using the Cell Counting Kit-8 (CCK-8) assay (Dojindo, Japan). The cells (1 × 104/well) were plated on 96-well plates for 24 h, followed by treatment with the gel-CM from the serum-free media (gel-CM), serum-deprivation media (2% FBS), and standard media (10% FBS). After treatments for 24, 48, and 72 h, the cell viability was measured using a CCK-8 assay kit (Dojindo, Japan) according to the manufacturer’s instructions, respectively. All experiments were performed three times.
5
Cytotoxicity Evaluation of Nanoparticles
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Cytotoxicity assays were prepared using both healthy and cancerous human cell lines: HDF (ATCC® PCS-201-012; Manassas, VA) and melanoma (ATCC CRL-1619; ATCC) cells. The cells were grown separately in DMEM (Thermo Fisher Scientific), 10% fetal bovine serum (FBS; ATCC 30-2020; ATCC) and 1% penicillin/streptomycin (Thermo Fisher Scientific). To quantify the cytotoxicity of the NPs toward the cell lines, cell viability (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium [MTS]) assays (CellTiter 96® AQueous One Solution Cell Proliferation Assay; Promega Corporation, Fitchburg, WI, USA) were carried out. The cells were seeded at a determined concentration of 5×104 cells/mL in 96-well plates and placed at 37°C in a humidified incubator with a 5% carbon dioxide (CO2) atmosphere for 24 hours; the media was then removed, and different concentrations of the NP colloids prepared in DMEM were added to each one of the wells. The plate was then incubated for 24 hours under the same environmental conditions. Following the second incubation period, the media was removed and MTS solution was added, leading to an incubation period of 4 hours before placing the plate in a spectrophotometer. The absorbance was then measured at 490 nm, and the cell viability in response to various NP concentrations was determined.
6
Fibroblast Cell Culture on Coated Glasses
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Human fibroblast (HDFa) cell culture (ATCC® PCS201 012™) was grown in T25 cell culture flask by using DMEM (high glucose 4.5 g/L, +glutamine, w/o pyruvate, Sigma®, USA) included 10% FBS (Sigma®, USA) and 1% Penicillin/streptomycin (10,000 IU/ml 10,000 µg/ml, Thermo Fisher Scientific®, USA) at 37 °C, 5% CO2 humidified incubator. After the culture reached the confluency, cells were detached from the flask surface via the use of Trypsin-EDTA (0.25%, Sigma®, USA) solution. The cell culture was seeded as 1 × 105 number of cells on the tungsten–germanium coated borosilicate glasses in 12-well plate and incubated for 24 h at 37 °C, 5% CO2 incubator to analyze attached cells. Experimental groups were classified as coated materials, uncoated borosilicate glasses, and negative control (cell culture plate surface), and each group was analyzed in triplicate samples.
7
Electrical Characterization of Fibroblast Cultures
8
Cytotoxicity and Photoprotection of (-)-Loliode
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HDF cells (ATCC®PCS201012™, Manassas, VA, USA) were cultured in the medium mixed with F-12 and DMEM (1:3) supplemented with 10% FBS and 1% P/S. HDF cells were seeded at a concentration of 5.0 × 104 cells/mL for experiments. To analyze the cytotoxicity of (-)-loliode on HDF cells, HDF cells were seeded and incubated with (-)-loliode (6.25, 12.5, 25, 50, and 100 μg/mL). After 24 h, the cell viabilities of (-)-loliode-treated HDF cells were measured by the MTT assay according to the method described by Wang et al. [23 (link)]. To evaluate the photoprotective effect of (-)-loliode in HDF cells, HDF cells were seeded and treated with (-)-loliode (6.25, 12.5, and 25 μg/mL). (-)-Loliode-treated cells were exposed to UVB (50 mJ/cm2) and the intracellular ROS level and the viability of UVB-irradiated HDF cells were determined with the DCF-DA assay and the MTT assay, respectively [11 (link)]. In addition, the collagen synthesis level and the expression of MMPs were assessed with ELISA kits (Sigma, St. Louis, MO, USA) using the cell culture medium [1 (link),11 (link)].
9
Evaluating AESIS-1 Cytotoxicity in HDFs
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Normal adult human dermal fibroblasts (HDFs) were obtained from ATCC (PCS-201-012, ATCC, Manassas, VA, USA) and cultured in Dulbecco’s modified Eagle’s medium (DMEM)/high glucose (Thermo Fisher Scientific, Waltham MA, USA) supplemented with 100 U/mL penicillin, 100 μg/mL streptomycin, and 10% fetal bovine serum (FBS, Thermo Fisher Scientific, Waltham MA, USA). The cells were frozen in liquid vapor nitrogen at −130 °C until use. Upon thawing, the cells were grown and maintained in a 5% CO2 incubator at 37 °C. Cell viability after 24 h of AESIS-1 treatment was measured with a Trypan blue exclusion assay [57 (link)]. The HDF cells (2 × 105 cells/6-well plate) were treated with AESIS-1 (1, 10, 100, 1000, 10,000 ng/mL) in serum-free media, and the unstained viable cell number was counted using a hemacytometer after 24 h of incubation.
10
Cell Line Cultivation and Maintenance
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Seven different cell lines, HeLa (ATCC® CCL-2™), HepG2 (ATCC® HB-8065™), MCF-7 (ATCC® HTB-22™), SW-620 (ATCC® CCL-227™), HCT116 (ATCC® CCL-247™), CCD 841 CoN (ATCC® CRL-1790™) and normal human dermal fibroblast (NHDF) (ATCC® PCS-201-012™) were used in this study. Each cell line was cultured in CO2 incubator at 37 °C and 5% CO2. Cells were cultured in DMEM supplemented with 10% fetal bovine serum (FBS), 60 mg/mL of penicillin and 100 mg/mL of streptomycin.
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