Caco2 (human, colon), Calu3 (human, lung), HUH7 (human, liver), VEROE6 (African green monkeys, kidney), and 293T (human, kidney) cell lines were grown in Dulbecco’s Modified Eagle Medium (DMEM) with Ultraglutamine (EuroClone, Italy) supplemented with 10% (or 20% for Calu3) Fetal Bovine Serum (FBS, EuroClone, Italy), 10 mM of HEPES (1 M) (EuroClone, Italy), 1% Penicillin-Streptomycin (100X) (EuroClone, Italy), 1% Non Essential Aminoacids (NEAA) (100X) (EuroClone, Italy), 1% NA Pyruvate (100 mM) (Euroclone, Italy); Raji (human B cell lymphoma) cells were grown in RPMI 1640 medium with Ultraglutamine (EuroClone, Italy) supplemented with 10% FBS, 10 mM of HEPES (1 M), 1% Penicillin-Streptomycin (100X), 1% NEAA (100X), 1% NA Pyruvate (100 mM). All cell lines were cultured in incubators set at 37 °C with 5% CO2 and passaged every 2–3 days.
Na pyruvate
Manufactured by Euroclone
Sourced in Italy
NA Pyruvate is a laboratory reagent used in various biochemical and cell culture applications. It serves as a source of pyruvate, a key intermediate in cellular metabolism. The product is typically supplied in a powder or solution form.
Automatically generated - may contain errors
Lab products found in correlation
Non essential amino acids (neaa), by Euroclone (2 mentions)
Fetal bovine serum (fbs), by Euroclone (2 mentions)
Ultraglutamine, by Euroclone (1 mentions)
Non essential amino acids neaa, by Euroclone (1 mentions)
Penicillin streptomycin, by Euroclone (1 mentions)
Hepes, by Euroclone (1 mentions)
Pen strep, by Euroclone (1 mentions)
Ccl 171, by Thermo Fisher Scientific (1 mentions)
Triton x 100, by Merck Group (1 mentions)
Dmem f12, by Thermo Fisher Scientific (1 mentions)
Hi fbs, by Thermo Fisher Scientific (1 mentions)
Minimum essential medium (mem), by Thermo Fisher Scientific (1 mentions)
Kanamycin, by Thermo Fisher Scientific (1 mentions)
Vr 740, by American Type Culture Collection (1 mentions)
3 4 5 dimethylthiazol 2 yl 2 5 diphenyl 2h tetrazolium bromide, by Merck Group (1 mentions)
Glutamine, by Euroclone (1 mentions)
Penicillin streptomycin, by Lonza (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Trypsin, by Lonza (1 mentions)
3 protocols using na pyruvate
1
Cell Culture Conditions for Diverse Cell Lines
2
Propagation and Infection Assay of hCoV-229E
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
hCoV229E (ATCC® VR-740™) was in propagated in MRC-5 cells (ATCC:CCL-171™) maintained in MEM (Gibco) supplemented with 10% v/v fetal beef serum (FBS HI; Gibco), 1 mM Na Pyruvate (Euroclone), 1 mM Non Essential Amino Acids (Euroclone) and 1x Pen-strep (Euroclone) and kept under 5% CO2 on 37 °C. BEAS-2B cells were kindly provided by Pierre-Olivier Vidalain were maintained in DMEM/F-12 (Gibco), 5% FBS HI (Gibco), 1% Kanamycin (Thermo-Fisher Scientifics), at 37 °C with 5% CO2. 2*104 cells per well were seeded in transparent 96 well plate and incubated over night in order to reach 90% confluency. 24 h later cells were infected with hCoV-229E m. o.i. 0.06 in DMEM/F-12 in presence of compound or 0,1% DMSO (untreated controls). The cells were incubated for 2 h at 35 °C with 5% CO2, then the virus was removed, replaced with complete medium with or without compound, and incubated at 35 °C with 5% CO2. After 72 h, 20 μL of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (Sigma-Aldrich) dissolved in PBS at 7,5 mg/mL were added to each well and incubated at 37 °C with 5% CO2 for 1 h. Then the supernatant was removed and cells were lysed with 100 μL/well of: 10% 2-Propanol, 0,004% Triton-X-100 (Sigma-Aldrich), 0,0004% HCl, then the absorbance was read at 570 nm with a plate reader Victor Nivo5 PerkinElmer.
3
HepG2 Cell Culture and Hydrogel Embedding
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Two
vials containing approximately 2 million HepG2 cells at passage 99
were kindly provided by “Istituto Zooprofilattico Sperimentale
della Lombardia e dell’ Emilia-Romagna”. Cells were
thawed, counted in a hemocytometer by means of the trypan blue (Sigma-Aldrich,
Lot. no. RNBD9396, US) exclusion assay, and seeded in a 6-well plate
with a seeding ratio of approximately 300,000 cells/well. The medium
used to culture cells was composed of EMEM with Earl’s salts
(EuroClone, Cat. no. ECB2071L, IT), 10% (v/v) FBS (EuroClone, Lot.
no. EU-S021179, IT), 1% (v/v) Na pyruvate (EuroClone, Lot. no. EU-M00QU,
IT), 1% (v/v) glutamine (EuroClone, Lot. no. EU-M0150017, IT) and
1% (v/v) penicillin–streptomycin (Lonza, Lot. no. 2MB027, BE),
and was refreshed every 48 h. Cells were expanded up to passage 6
and then cryopreserved (106 cells/cryovial) in complete
medium + 10% (v/v) DMSO (Sigma-Aldrich, Lot. no. SZBD2870V, US). To
be embedded within hydrogels, cells were expanded, detached with trypsin
(Lonza, Lot. no. 4MB146, BE), counted, suspended in a fresh medium,
and embedded within gels as described in 2.3 at a final density of
3 × 106 cells/gel. Hydrogels were then submerged with
2 mL of fresh medium and incubated at 37 °C, 95% humidity, and
5% CO2.
vials containing approximately 2 million HepG2 cells at passage 99
were kindly provided by “Istituto Zooprofilattico Sperimentale
della Lombardia e dell’ Emilia-Romagna”. Cells were
thawed, counted in a hemocytometer by means of the trypan blue (Sigma-Aldrich,
Lot. no. RNBD9396, US) exclusion assay, and seeded in a 6-well plate
with a seeding ratio of approximately 300,000 cells/well. The medium
used to culture cells was composed of EMEM with Earl’s salts
(EuroClone, Cat. no. ECB2071L, IT), 10% (v/v) FBS (EuroClone, Lot.
no. EU-S021179, IT), 1% (v/v) Na pyruvate (EuroClone, Lot. no. EU-M00QU,
IT), 1% (v/v) glutamine (EuroClone, Lot. no. EU-M0150017, IT) and
1% (v/v) penicillin–streptomycin (Lonza, Lot. no. 2MB027, BE),
and was refreshed every 48 h. Cells were expanded up to passage 6
and then cryopreserved (106 cells/cryovial) in complete
medium + 10% (v/v) DMSO (Sigma-Aldrich, Lot. no. SZBD2870V, US). To
be embedded within hydrogels, cells were expanded, detached with trypsin
(Lonza, Lot. no. 4MB146, BE), counted, suspended in a fresh medium,
and embedded within gels as described in 2.3 at a final density of
3 × 106 cells/gel. Hydrogels were then submerged with
2 mL of fresh medium and incubated at 37 °C, 95% humidity, and
5% CO2.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!