Scf cytokines

Manufactured by Thermo Fisher Scientific

SCF cytokines are proteins produced by cells that play a role in cell signaling and cellular processes. They are used in various research and analytical applications in the life sciences industry.

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3 protocols using scf cytokines

Maintaining Diverse Leukemia Cell Lines

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U937, HL-60, and 293T cells were purchased from ATCC. Ba/F3 cells were purchased from DSMZ. THP-1, MV4-11, and MOLM-14 cells were provided by Scott Armstrong. MOLM-13 cells were provided by Benjamin Ebert. All cell lines were maintained in RPMI 1640 (Cellgro) supplemented with 1% penicillin/streptomycin (PS; Cellgro) and 10% FBS (Sigma-Aldrich) at 37°C with 5% CO₂. The 293T cells were maintained in Dulbecco’s modified Eagle’s medium (Invitrogen) supplemented with 10% FBS (Invitrogen) and 1% PS. Ba/F3 cells were maintained in RPMI-1640 supplemented with 1% PS, 10% FBS, and IL-3 (PeproTech).
Primary patient AML blasts were collected from peripheral blood or bone marrow aspirate after obtaining informed consent under Dana-Farber Institute Internal Review Board–approved protocols. Mononuclear cells were isolated using Ficoll-Paque Plus (GE Healthcare), and red blood cells were lysed (eBioscience). AML blasts from patients were maintained in SFEM (Stemcell Technologies) with SCF, FLT3L, IL-3, IL-6, and GCSF (PeproTech). For drug treatment of primary AML cells, RPMI-1640 with PS and FBS were used.
Murine MLL-AF9–expressing leukemia cells were obtained from a quaternary bone marrow transplant model from Benjamin Ebert’s laboratory. Cells were maintained in SFEM media (Stemcell Technologies) with murine IL-3, IL-6, and SCF cytokines (PeproTech).

Pikman Y., Puissant A., Alexe G., Furman A., Chen L.M., Frumm S.M., Ross L., Fenouille N., Bassil C.F., Lewis C.A., Ramos A., Gould J., Stone R.M., DeAngelo D.J., Galinsky I., Clish C.B., Kung A.L., Hemann M.T., Vander Heiden M.G., Banerji V, & Stegmaier K. (2016). Targeting MTHFD2 in acute myeloid leukemia. The Journal of Experimental Medicine, 213(7), 1285-1306.

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Cryopreservation and Culture of Primary Hematopoietic Cells

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Primary cells were cultured as previously done in Pei et al.^{66 (link)}. Primary human AML cells, normal cord/peripheral blood mononuclear cells (CBMCs/PBMCs) were resuspended at about 100–200 e⁶ cells per ml in freezing media composed of 50% FBS (GE Healthcare), 10% DMSO (Sigma) and 40% IMDM media (Gibco) and then cryo-preserved in liquid nitrogen. Cells were thawed in 37 °C water bath, washed twice in thawing media composed of IMDM (Gibco), 2.5% FBS (GE Healthcare) and 10 ug/ml DNAse (Sigma). Normal CD34+ CBMCs/PBMCs were enriched from CBMCs/PBMCs using the CD34 MicroBead kit (Miltenyi Biotec). Cells were cultured in normal or complete serum-free media (SFM) in 37 °C, 5% CO₂ incubator. Normal SFM is composed of IMDM (Gibco), 20% BIT 9500 (STEMCELL Technologies), 10ug/ml LDL (Low Density Lipoprotein, Millipore), 55uM 2-Mercaptoethanol (Gibco) and 1% Pen/Strep (Gibco). Complete SFM were made by supplementing the normal SFM with FLT-3, IL-3 and SCF cytokines (PeproTech), each at 10 ng/ml. MDS-L cells were a generous gift from Daniel Starczynowski and grown in RPMI-1640 medium with 10% FBS, 1% penicillin-streptomycin and supplemented with 10 ng/ml IL-3^{67 (link)}.

Stevens B.M., Khan N., D’Alessandro A., Nemkov T., Winters A., Jones C.L., Zhang W., Pollyea D.A, & Jordan C.T. (2018). Characterization and targeting of malignant stem cells in patients with advanced myelodysplastic syndromes. Nature Communications, 9, 3694.

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Cryopreservation and culture of AML samples

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Primary human AML specimens and normal MPB samples were resuspended at about 100-200 e6 cells per ml in freezing media composed of 50% FBS (GE Healthcare), 10% DMSO (Sigma) and 40% IMDM media (GIBCO) and then cryo-preserved in liquid nitrogen. Cells were thawed in 37°C water bath, washed twice in thawing media composed of IMDM (GIBCO), 2.5% FBS (GE Healthcare) and 10 ug/ml DNase (Sigma). Normal CD34+ HSPCs were enriched from thawed MPB samples using the CD34 MicroBead kit (Miltenyi Biotec). Cells were cultured in complete serum-free media (SFM) in 37°C, 5% CO2 incubator. SFM is composed of IMDM (GIBCO), 20% BIT 9500 (STEMCELL Technologies), 10ug/ml LDL (Low Density Lipoprotein, Millipore), 55uM 2-Mercaptoethanol (GIBCO) and 1% Pen/Strep (GIBCO). Complete SFM were made by supplementing the SFM with FLT-3, IL-3 and SCF cytokines (PeproTech), each at 10 ng/ml.

Kruglyak S., Durrett R.T., Schug M.D, & Aquadro C.F. (1998). Equilibrium distributions of microsatellite repeat length resulting from a balance between slippage events and point mutations. Proceedings of the National Academy of Sciences of the United States of America, 95(18), 10774-10778.

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