Log In Sign up
The largest database of trusted experimental protocols

Defined fbs

Manufactured by Thermo Fisher Scientific
Visit Supplier

Defined FBS is a high-quality serum product specifically formulated for cell culture applications. It provides a consistent and defined nutrient source to support cell growth and proliferation.

Automatically generated - may contain errors

Lab products found in correlation

Non essential amino acids (neaa), by Thermo Fisher Scientific (3 mentions) Activin a, by Cell Guidance Systems (2 mentions) Non essential amino acids (neaa), by R&D Systems (2 mentions) Noggin, by R&D Systems (2 mentions) Rpmi 1640 media, by Thermo Fisher Scientific (2 mentions) Bone morphogenetic protein 4 (bmp4), by R&D Systems (2 mentions) Retinoic acid, by Merck Group (2 mentions) Accutase, by STEMCELL (2 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (2 mentions) Hgm csf, by Thermo Fisher Scientific (1 mentions) Hil 3, by Thermo Fisher Scientific (1 mentions) Af2420, by R&D Systems (1 mentions) Hm csf, by Thermo Fisher Scientific (1 mentions) E myco plus kit, by iNtRON Biotechnology (1 mentions) Puromycin, by Merck Group (1 mentions) Penicillin, by Thermo Fisher Scientific (1 mentions) Streptomycin, by Thermo Fisher Scientific (1 mentions) Trypsin, by Thermo Fisher Scientific (1 mentions) Dulbecco s modified eagle s medium dmem, by Thermo Fisher Scientific (1 mentions)

6 protocols using defined fbs

1

Directed Differentiation of Gastric Organoids

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
The protocol for directed differentiation of gastric organoids was adapted from our previous protocol7 (link), and Supplementary Table 1 contains the complete list of media and growth factors for each stage. For differentiation, hPSCs were dissociated into single cells using Accutase (Stem Cell Technologies) and plated into 24-well plates at a density of roughly 200,000 cells per well in mTesR1 with Y-27632 (10 μM; Stemgent). The following day, cells were differentiated into definitive endoderm (DE) by adding Activin A (100 ng/ml; Cell Guidance Systems) in RPMI 1640 media (Invitrogen) for three days. Media was also supplemented with NEAA (1×; Gibco) and defined FBS (dFBS; Invitrogen) at 0%, 0.2%, and 2.0% on days 1, 2, and 3, respectively. Additionally, BMP4 (50 ng/ml; R&D Systems) was added on the first day. Subsequently, DE was differentiated to posterior foregut endoderm by exposing cells to CHIR99021 (2 μM; Stemgent), FGF4 (500 ng/ml; R&D Systems), and Noggin (200 ng/ml; R&D systems) for three days in RPMI 1640 supplemented with NEAA and 2.0% dFBS. Retinoic acid (2 μM; Sigma Aldrich) was added for the final day. Media was changed every day. This process resulted in the spontaneous formation of three-dimensional posterior foregut spheroids.
McCracken K.W., Zhang X, & Wells J.M. (2017). Wnt/β-catenin promotes gastric fundus specification in mice and humans. Nature, 541(7636), 182-187.
+ Open protocol
+ Expand
2

Directed Differentiation of Gastric Organoids

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
The protocol for directed differentiation of gastric organoids was adapted from our previous protocol7 (link), and Supplementary Table 1 contains the complete list of media and growth factors for each stage. For differentiation, hPSCs were dissociated into single cells using Accutase (Stem Cell Technologies) and plated into 24-well plates at a density of roughly 200,000 cells per well in mTesR1 with Y-27632 (10 μM; Stemgent). The following day, cells were differentiated into definitive endoderm (DE) by adding Activin A (100 ng/ml; Cell Guidance Systems) in RPMI 1640 media (Invitrogen) for three days. Media was also supplemented with NEAA (1×; Gibco) and defined FBS (dFBS; Invitrogen) at 0%, 0.2%, and 2.0% on days 1, 2, and 3, respectively. Additionally, BMP4 (50 ng/ml; R&D Systems) was added on the first day. Subsequently, DE was differentiated to posterior foregut endoderm by exposing cells to CHIR99021 (2 μM; Stemgent), FGF4 (500 ng/ml; R&D Systems), and Noggin (200 ng/ml; R&D systems) for three days in RPMI 1640 supplemented with NEAA and 2.0% dFBS. Retinoic acid (2 μM; Sigma Aldrich) was added for the final day. Media was changed every day. This process resulted in the spontaneous formation of three-dimensional posterior foregut spheroids.
McCracken K.W., Zhang X, & Wells J.M. (2017). Wnt/β-catenin promotes gastric fundus specification in mice and humans. Nature, 541(7636), 182-187.
+ Open protocol
+ Expand
3

iPSC-Derived Microglial Characterization

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
iPSCs were first differentiated into hematopoietic progenitor cells following manufacturer’s instructions using a commercially available kit (StemCell Technologies #05310) as described before (Andreone et al., 2020 (link)). HPCs positive for identity markers CD34, CD43, and CD45 were transferred to a plate containing primary human astrocytes and co-cultured using media C adapted from a previous study (Pandya et al., 2017 (link)). Once floating cells in co-culture are predominantly (>80%) mature microglia, cells were plated for 3–4 days prior to experiments. Full characterization of human iPSC-derived microglia and additional details on the differentiation protocol has been published elsewhere (Andreone et al., 2020 (link)). All the experiments using GRN+/+ and GRN−/− iMG were performed in IMDM (Gibco) media supplemented with 10% defined FBS (Gibco), 1% Penicillin/Streptomycin (Gibco), 20 ng/mL of hIL3 (Peprotech), 20 ng/mL of hGM-CSF (Peprotech) and 20 ng/mL of hM-CSF (Peprotech) (referred to as “C+++ Media”). Immunostainings of PGRN were conducted using the anti-progranulin goat polyclonal antiserum (R&D Systems # AF2420, 1:250) to verify the absence of immunoreactivity in knockout iMG.
Logan T., Simon M.J., Rana A., Cherf G.M., Srivastava A., Davis S.S., Yoon Low R.L., Chiu C.L., Fang M., Huang F., Bhalla A., Llapashtica C., Prorok R., Pizzo M.E., Calvert M.E., Sun E.W., Hsiao-Nakamoto J., Rajendra Y., Lexa K.W., Srivastava D.B., van Lengerich B., Wang J., Robles-Colmenares Y., Kim D.J., Duque J., Lenser M., Earr T.K., Nguyen H., Chau R., Tsogtbaatar B., Ravi R., Skuja L.L., Solanoy H., Rosen H.J., Boeve B.F., Boxer A.L., Heuer H.W., Dennis M.S., Kariolis M.S., Monroe K.M., Przybyla L., Sanchez P.E., Meisner R., Diaz D., Henne K.R., Watts R.J., Henry A.G., Gunasekaran K., Astarita G., Suh J.H., Lewcock J.W., DeVos S.L, & Di Paolo G. (2021). Rescue of a lysosomal storage disorder caused by Grn loss-of-function with a brain penetrant progranulin biologic. Cell, 184(18), 4651-4668.e25.
+ Open protocol
+ Expand
4

RAW264.7 macrophages and T. gondii protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols
RAW264.7 macrophages (ATCC TIB-17) were maintained in DMEM (Gibco)
containing 10% defined FBS (Gibco) at 37°C, 5%
CO2 and passaged by dilution upon reaching confluency. T.
gondii RH strain parasites were maintained by passage in HFFs,
cultured in DMEM 3% FBS, as previously described 26 . The genotypes of parasites used in
recruitment assays were
RHΔku80ΔhxgprtΔuprt::CBR(herein described as wild type) and
RHΔku80Δrop18::HXGPRTΔuprt::CBR(herein described as Δrop18 mutant), as described
previously 58 . All cultures
were determined to be mycoplasma negative using the e-Myco plus kit (Intron
Biotechnology).
Simpson C., Jones N.G., Hull-Ryde E.A., Kireev D., Stashko M., Tang K., Janetka J., Wildman S.A., Zuercher W.J., Schapira M., Hui R., Janzen W, & Sibley L.D. (2015). Identification of small molecule inhibitors that block the Toxoplasma gondii rhoptry kinase ROP18. ACS infectious diseases, 2(3), 194-206.
+ Open protocol
+ Expand
5

Establishing RIPK3-HA and CK1γ Knockout Cell Lines

Check if the same lab product or an alternative is used in the 5 most similar protocols
HT-29 cells were cultured in McCoy’s 5A medium (Hyclone) and other cells were in Dulbecco’s Modified Eagles Medium (DMEM; Hyclone), supplemented with 10% defined FBS (Gibco) and 100 U/mL penicillin. Cells were maintained in a 37 °C incubator at 5% CO2. HeLa cells were transfected with pcDNA3 encoding RIPK3-HA using iN-fect reagents (iNtRON Biotechnology) and selected with G418 (Sigma-Aldrich) to establish the HeLa/RIPK3-HA cell line. CK1γ knockout cells were generated using CRISPR/Cas9 system with LentiCRISPR (pXPR_001) that expresses each gene-targeting sgRNA. HeLa/RIPK-HA cells were transduced with the lentiviral delivery and selected with puromycin (Sigma-Aldrich). For the controls, non-specific sgRNA was transduced into cells with the same procedure.
Lee S.Y., Kim H., Li C.M., Kang J., Najafov A., Jung M., Kang S., Wang S., Yuan J, & Jung Y.K. (2019). Casein kinase-1γ1 and 3 stimulate tumor necrosis factor-induced necroptosis through RIPK3. Cell Death & Disease, 10(12), 923.
+ Open protocol
+ Expand
6

Murine Pluripotent Stem Cell Culture

Check if the same lab product or an alternative is used in the 5 most similar protocols
The cells were cultured as described previously [9] (link). In brief, STO feeder cells were used for following two mouse ES cell-lines. C57BL/6-background mouse ES cells (mESCs, accession #. SCRC-1002; ATCC), and 129/Ola strain-derived mouse ES cells (provided by Jeong Mook Lim, Seoul National University, Seoul, Korea). Protein-based-iPSCs were generated from mouse cardiac fibroblast (isolated from heart of C57BL/6 mice) and skin fibroblast (harvested from dermis of FVB mice) using ES cell-derived protein extracts (Protein-iPSCs). STO cells were cultured in Dulbecco's modified Eagle's medium (DMEM; GIBCO, Grand Island, NY) high glucose supplemented with 10% Fetal bovine Serum (FBS; GIBCO), and 100 U/ml Penicillin streptomycin (GIBCO). One day before subculturing mESCs or P-iPSCs, STO cells were treated with Mitomycin C (10 μg/ml medium, sigma-Aldrich, St Louis, MO) and seeded on a new 0.1% gelatin-coated dish. Propagating mESCs and P-iPSCs were cultured in DMEM (GIBCO) with 10% defined FBS (GIBCO), 2 mM L-glutamine (GIBCO), 1X NEAA (GIBCO), 1 mM 2-Mercaptoethanol (Sigma-Aldrich, St Louis, MO), and 100 units/ml Penicillin/100 μg/ml streptomycin (GIBCO) (ES media). In ES media, 2000 U/ml of ESGRO® LIF (leukemia inhibitory factor, Chemicon) was added to maintain pluripotency. mESCs and P-iPSCs were dissociated with 0.05% trypsin (GIBCO) and passaged on STO every 2∼3 days.
Kwon Y.W., Chung Y.J., Kim J., Lee H.J., Park J., Roh T.Y., Cho H.J., Yoon C.H., Koo B.K, & Kim H.S. (2014). Comparative Study of Efficacy of Dopaminergic Neuron Differentiation between Embryonic Stem Cell and Protein-Based Induced Pluripotent Stem Cell. PLoS ONE, 9(1), e85736.
+ Open protocol
+ Expand

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

Sign up now

Revolutionizing how scientists
search and build protocols!