LLC-PK1 cells (porcine kidney cells) were purchased from the ATCC (ATCC CL-101) and cultured in Dulbecco's modified Eagle's medium (Invitrogen, USA) supplemented with 10% fetal bovine serum at 37 °C in a humidified 5% CO2 incubator. HEK-293T cells, obtained from the China Center for Type Culture Collection (CCTCC), were cultured in Dulbecco's modified Eagle's medium (Invitrogen, USA) supplemented with 10% fetal bovine serum under the same conditions described above. PDCoV strain CHN-HN-2014 (GenBank accession number KT336560) was isolated from a piglet with acute diarrhea in China in 2014 (Dong et al., 2016 (link)). Sendai virus (SEV) was obtained from the Centre of Virus Resource and Information, Wuhan Institute of Virology, Chinese Academy of Sciences. Mouse monoclonal antibodies (MAbs) against hemagglutinin (HA) and β-actin were purchased from Medical and Biological Laboratories (Japan). Rabbit polyclonal antibodies directed against IRF3, phosphorylated IRF3 (p-IRF3), p65, and phosphorylated p65 (p-p65) were purchased from ABclonal (China) and Cell Signaling Technology.
Atcc cl 101
Manufactured by American Type Culture Collection
Sourced in United States
The ATCC CL-101 is a laboratory equipment product offered by American Type Culture Collection. It serves as a core function for cell culture applications. The detailed technical specifications and intended use of this product are not available in this concise and unbiased format.
Automatically generated - may contain errors
Lab products found in correlation
Antibiotic antimycotic, by Thermo Fisher Scientific (2 mentions)
Dulbecco s modified eagle s medium, by Thermo Fisher Scientific (1 mentions)
Hek293t cell, by National Collection of Authenticated Cell Cultures (1 mentions)
Mem non essential amino acids neaas, by Thermo Fisher Scientific (1 mentions)
Hepes, by Thermo Fisher Scientific (1 mentions)
Minimum essential medium (mem), by Thermo Fisher Scientific (1 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Non essential amino acids (neaa), by Thermo Fisher Scientific (1 mentions)
4 protocols using atcc cl 101
1
Porcine Kidney Cell-Based Antiviral Protocol
2
Porcine Kidney Cell Culture for PDCoV Isolation
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LLC porcine kidney
(LLC-PK) cells were purchased from the ATCC (ATCC CL-101) and cultured
at 37 °C with 5% CO2 in the modified Eagle medium
(MEM) (Invitrogen, Carlsbad, USA) containing 10% heat-inactivated
fetal bovine serum (FBS) (Gibco, Grand Island, NY, USA), 1% MEM nonessential
amino acids (NEAAs) (Gibco, Grand Island, NY, USA), 1% HEPES (Gibco,
Grand Island, NY, USA), and 1% antibiotic-antimycotic (Gibco, Grand
Island, NY, USA). The PDCoV strain CH/XJYN/2016 (GenBank accession
number: MN064712) was isolated from intestinal samples of a suckling
piglet experiencing acute diarrhea using LLC-PK cells in our laboratory.
(LLC-PK) cells were purchased from the ATCC (ATCC CL-101) and cultured
at 37 °C with 5% CO2 in the modified Eagle medium
(MEM) (Invitrogen, Carlsbad, USA) containing 10% heat-inactivated
fetal bovine serum (FBS) (Gibco, Grand Island, NY, USA), 1% MEM nonessential
amino acids (NEAAs) (Gibco, Grand Island, NY, USA), 1% HEPES (Gibco,
Grand Island, NY, USA), and 1% antibiotic-antimycotic (Gibco, Grand
Island, NY, USA). The PDCoV strain CH/XJYN/2016 (GenBank accession
number: MN064712) was isolated from intestinal samples of a suckling
piglet experiencing acute diarrhea using LLC-PK cells in our laboratory.
3
Porcine Deltacoronavirus Infection Assay
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LLC-PK1 cells (ATCC CL-101) from ATCC (the American Type Culture Collection) was cultured in Dulbecco’s modified Eagle’s medium (DMEM) (Gibco, Gaithersburg, MD, USA) supplemented with 1% antibiotic-antimycotic (Gibco, USA), 1% nonessential amino acids solution (NEAA) (Gibco, USA), and 10% fetal bovine serum (FBS) (Gibco, Fitzroy North, Victoria, Australia). The PDCoV CHN-HN-1601 strain (GenBank accession no. MG832584 ) was provided by Hanchun Yang, China Agricultural University, Beijing, China. The PDCoV strain was used to infect LLC-PK1 cells at an MOI of 0.1 or 1. After incubation for 1 h at 37°C in a 5% CO2 incubator, cells were washed three times to remove the unbound viruses and cultured in maintenance medium (DMEM supplemented with 0.4% trypsin) at 37°C.
4
Porcine Proximal Tubule Cell Differentiation Assay
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LLC PK-1 (Lilly Laboratories Porcine Kidney-1) porcine proximal tubule cells (ATCC-CL-101, ATCC, Manassas, VA) were expanded in 75 cm2 flasks at 37°C with 5% CO2 and passed once confluence was reached. Culture medium consisted in a 1:1 Dulbecco’s Modified Eagle’s Medium-Ham’s F12 mix supplemented with 5% FBS, 15 mM HEPES, 0.1% Sodium bicarbonate, 100 UI.ml−1 Penicillin/Streptomycin and 50 nM Sodium selenite. LLC PK-1 cells were cultured between passage 7 and passage 25.
LLC PK-1 were seeded in four 60 mm Petri dishes (one per condition) and expanded up to sub-confluence in the routine cell culture medium.
Seeded LLC PK-1 sustained serum starvation and were fed with hormonally defined (25 µg.ml−1 insulin, 11 µg.ml−1 transferrin, 50 nM triiodothyronine, 0.1 µM dexamethasone, 0.1 µg.ml−1 desmopressin) fresh medium to engage epithelial differentiation, for 24 h.
Differentiated LLC PK-1 cells were exposed for 24 h to four different conditions: Control (vehicle: 96% Ethanol), 5 µM CsA, 0.05 µM Tac or 1 µM VIVIT (a specific Nuclear Factor of Activated T cells [NFAT] inhibitor [18 (link)]).
LLC PK-1 were seeded in four 60 mm Petri dishes (one per condition) and expanded up to sub-confluence in the routine cell culture medium.
Seeded LLC PK-1 sustained serum starvation and were fed with hormonally defined (25 µg.ml−1 insulin, 11 µg.ml−1 transferrin, 50 nM triiodothyronine, 0.1 µM dexamethasone, 0.1 µg.ml−1 desmopressin) fresh medium to engage epithelial differentiation, for 24 h.
Differentiated LLC PK-1 cells were exposed for 24 h to four different conditions: Control (vehicle: 96% Ethanol), 5 µM CsA, 0.05 µM Tac or 1 µM VIVIT (a specific Nuclear Factor of Activated T cells [NFAT] inhibitor [18 (link)]).
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