All cells were cultured at 37°C under 5% CO2 in serum-free defined media containing DMEM, transferrin (100 μg/mL), BSA (100μg/mL), putrescine (16 μg/mL), progesterone (60 ng/mL), sodium selenite (40 ng/mL), N-acetyl-L-cysteine (5 μg/mL), d-biotin (10 ng/mL), forskolin (4.2 μg/mL), insulin (5 μg/mL) (all from Sigma), glutamine (2 mM), sodium pyruvate (1 mM), B27 (2%), penicillin–streptomycin (100 U/mL) (all from Invitrogen), Trace Elements B (1x, Cellgro), and LIF (20 ng/mL, Chemicon). Proliferation medium also contained OPC mitogens PDGF-AA (20 ng/mL) and NT-3 (1 ng/mL) (both from PeproTech), while differentiation medium contained triiodothyronine (40 ng/mL, Sigma) without OPC mitogens. For experiments in which only mature OLGs were examined (and not any intervening stages of differentiation), the differentiation medium was modified by using DMEM/F12 base media (Invitrogen) and a higher dose of insulin (25 μg/mL) to maximize OLG survival.
Trace elements b
Manufactured by Corning
Trace Elements B is a laboratory solution that provides a source of essential trace elements required for cell culture and microbial growth. The solution contains a balanced mixture of micronutrients including iron, copper, manganese, zinc, cobalt, and molybdenum.
Automatically generated - may contain errors
Lab products found in correlation
D biotin, by Merck Group (4 mentions)
Sodium pyruvate, by Thermo Fisher Scientific (3 mentions)
Insulin, by Merck Group (3 mentions)
Triiodothyronine, by Merck Group (3 mentions)
Forskolin, by Merck Group (3 mentions)
Putrescine, by Merck Group (2 mentions)
Pdgf aa, by Thermo Fisher Scientific (2 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions)
Bovine serum albumin (bsa), by Merck Group (2 mentions)
Dulbecco modified eagle medium (dmem), by Merck Group (2 mentions)
Insulin, by Thermo Fisher Scientific (2 mentions)
Glutamine, by Merck Group (2 mentions)
Sodium selenite, by Merck Group (2 mentions)
Transferrin, by Merck Group (2 mentions)
Progesterone, by Merck Group (2 mentions)
N acetyl l cysteine, by Merck Group (2 mentions)
Dmem f12 base media, by Thermo Fisher Scientific (2 mentions)
B27 supplement, by Thermo Fisher Scientific (2 mentions)
N acetylcysteine, by Merck Group (2 mentions)
Dnase 1, by Roche (1 mentions)
6 protocols using trace elements b
1
Oligodendrocyte Differentiation Protocol
2
Oligodendrocyte Differentiation Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
All cells were cultured at 37°C under 5% CO2 in serum-free defined media containing DMEM, transferrin (100 μg/mL), BSA (100μg/mL), putrescine (16 μg/mL), progesterone (60 ng/mL), sodium selenite (40 ng/mL), N-acetyl-L-cysteine (5 μg/mL), d-biotin (10 ng/mL), forskolin (4.2 μg/mL), insulin (5 μg/mL) (all from Sigma), glutamine (2 mM), sodium pyruvate (1 mM), B27 (2%), penicillin–streptomycin (100 U/mL) (all from Invitrogen), Trace Elements B (1x, Cellgro), and LIF (20 ng/mL, Chemicon). Proliferation medium also contained OPC mitogens PDGF-AA (20 ng/mL) and NT-3 (1 ng/mL) (both from PeproTech), while differentiation medium contained triiodothyronine (40 ng/mL, Sigma) without OPC mitogens. For experiments in which only mature OLGs were examined (and not any intervening stages of differentiation), the differentiation medium was modified by using DMEM/F12 base media (Invitrogen) and a higher dose of insulin (25 μg/mL) to maximize OLG survival.
3
Isolation and Culture of Oligodendrocytes
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mice (CD-1) cortices of P1/2 pups were isolated, diced, and digested in a solution containing papain (1.54 µg/ml; Worthington), l -cysteine (360 µg/ml; Sigma-Aldrich), and DNaseI (703 µg/ml; Roche) for 25 min at 37°C as outlined previously (O’Meara et al., 2011 (link)). Dissociated cells were plated at high density on T75 tissue culture flasks coated with poly-l -lysine (PLL; Sigma-Aldrich) for 7–10 d in high-glucose DMEM supplemented with 1 mM sodium pyruvate, 2 mM l -glutamine or 1× GlutaMAX, 50 U/ml penicillin, 50 µg/ml streptomycin, and 10% FBS. Confluent T75 flasks were shaken overnight 18–20 h at 220 rpm, and the supernatant was incubated for 30 min on a tissue culture plate to remove adherent cells. Nonadherent cells were plated at 5,000 cells per well in PLL-coated 96-well tissue culture plates in oligodendrocyte growth medium composed of high-glucose DMEM supplemented with 1 mM sodium pyruvate, 1× GlutaMAX, 50 U/ml penicillin, 50 µg/ml streptomycin, 1× B27 supplement (Thermo Fisher Scientific), 50 µg/ml holo-transferrin, 100 µg/ml albumin from bovine serum, 5 ng/ml sodium selenite, 16 µg/ml putrescine, 60 ng/ml progesterone, 400 ng/ml 3,3′,5-thiiodo-l -thryonine, 5 µg/ml bovine insulin, 5 µg/ml N-acetyl cysteine, 10 ng/ml d -biotin (Sigma-Aldrich), and 1× Trace Elements B (Corning). After 24 h in vitro, cells were treated with toxins for a defined period of time.
4
Isolation and Culture of Mouse Oligodendrocyte Progenitor Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mouse OPCs were isolated from the cortices of pups at postnatal days 3–8 as described previously.[30 ] Briefly, cortical tissues were dispersed into single cells and the cell suspension was then subjected to immunopanning with antibodies against GalC and O4 sequentially. The enriched Galc‐negative O4‐positive OPCs were plated into poly‐D‐lysine coated dishes and cultured with Mouse OPC growth medium (DMEM/F‐12 (GIBCO, Cat# 11330‐032) supplemented with 1% N2 supplement (GIBCO, Cat# A1370701), 2% B27 supplement (GIBCO Cat# A3582801), penicillin–streptomycin solution (MP Biomedicals, Cat# 0916700), 1% sodium pyruvate (GIBCO, Cat# 11360070), 1% L‐glutamine (Hyclone, Cat# SH30034), 10 ng mL−1 platelet‐derived growth factor‐aa (Peprotech, Cat#100‐13A), 10 ng mL−1 ciliary neurotrophic factor (Peprotech, Cat# 450‐13), 20 ng mL−1 human basic fibroblast growth factor (Sino Biological, Cat# 10014HNAE), 0.5 mg mL−1 insulin (Sigma, Cat# 91077), 5 mg mL−1N‐acetyl cysteine (Sigma, Cat# A8199), 10 ng mL−1 D‐biotin (Sigma, Cat# B4639), 5 mm forskolin (Sigma, Cat# F3917), and 0.1% Trace Elements B (Corning, Cat# 25‐022‐CI)). OPC differentiation medium contained the same components as growth medium except human basic fibroblast growth factor and platelet‐derived growth factor‐aa, but supplemented with 40 ng mL−1 triiodo‐thyronine (Sigma, Cat# T2877).
5
Murine Mesothelioma and Healthy Mesothelial Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
AB22 murine mesothelioma and MeT-5A human healthy mesothelial cell lines were purchased from Sigma and ATCC respectively. AB22 cells were cultured in RPMI media supplemented with 25 mM Hepes, 10% FBS, 1% P/S and 2 mM of Glutamine, while MeT-5A were cultured in Medium 199 supplemented with 10% FBS, 1% P/S, 8.7E−4 mM of bovine insulin, 3.3E−6 mM of human EGF, 4E−4 mM of hydrocortisone and Trace Elements B (Corning) and maintained in a 5% CO2 incubator at 37 °C.
6
Neural Progenitor Culture and Differentiation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The attached neural progenitors were collected and cultured in suspension from day 15 to day 20 which the culture medium was changed every other day with OIM which consisted of DMEM-F12 with 1:200 N2 supplement, 1:100 B27 supplement lacking vitamin A, 1 μM SAG (Millipore), 10 ng/mL hPDGF-AA (Peprotech), 10 ng/mL NT3 (Peprotech), 10 ng/mL IGF-I (Peprotech), 200 μM AA (Sigma), 1:1,000 Trace Elements B (Corning), 10 ng/mL T3 (Sigma). Then, the neural spheres were cultured with DM medium consisting of DMEM-F12 with N2 supplement (1:200), B27 supplement without vitamin A (1:100), 60 ng/mL T3, 10 ng/mL NT3, 10 ng/mL IGF-I, 200 μM AA, Trace Elements B (1:1,000), biotin (sigma) and 100 μM cAMP (Sigma). After 7–10 day of differentiation, cells were dissociated with accutase and reseeded at densities of 3 × 104 cells on Matrigel coated glass coverslips of 24-well plates for staining analysis.
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!