The human AML cell lines HL-60, Kasumi-1, and KG-1 were purchased from Procell (Wuhan, China). HL-60 and KG-1 cells were incubated in Iscove’s Modified Dulbecco Medium (IMDM, Procell) supplemented with 20% fetal bovine serum (FBS, HyClone, Utah, USA) and Kasumi-1 cells were cultured in Roswell Park Memorial Institute (RPMI)-1640 medium (Procell) supplemented with 20% FBS (HyClone). All the media were placed in an incubator at 37 °C and 5% CO2.
Iscove modified dulbecco media (imdm)
Manufactured by Procell
Sourced in China
IMDM is a cell culture medium commonly used for the growth and maintenance of various cell types in laboratory settings. It provides a balance of nutrients, vitamins, and other essential components required for cell proliferation and survival.
Automatically generated - may contain errors
Lab products found in correlation
Hl 60, by Procell (2 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (2 mentions)
Penicillin streptomycin solution, by Procell (2 mentions)
Mccoy 5a, by Procell (2 mentions)
Hct116, by Procell (2 mentions)
Hitransg a, by Genechem (2 mentions)
Rpmi 1640 medium, by Procell (1 mentions)
Kasumi 1, by Procell (1 mentions)
B 27 minus insulin, by Thermo Fisher Scientific (1 mentions)
Rpmi 1640, by Thermo Fisher Scientific (1 mentions)
Insulin transferrin selenium its, by Merck Group (1 mentions)
Glutamax supplement, by Thermo Fisher Scientific (1 mentions)
Basic fibroblast growth factor (bfgf), by Thermo Fisher Scientific (1 mentions)
Dexamethasone (dex), by Merck Group (1 mentions)
Hepatocyte growth factor (hgf), by Thermo Fisher Scientific (1 mentions)
Knockout serum replacement (ksr), by Thermo Fisher Scientific (1 mentions)
Iscove modified dulbecco media (imdm), by Thermo Fisher Scientific (1 mentions)
Activin a, by Thermo Fisher Scientific (1 mentions)
Hepatocyte culture medium, by Lonza (1 mentions)
β mercaptoethanol β me, by Merck Group (1 mentions)
8 protocols using iscove modified dulbecco media (imdm)
1
Culturing Human AML Cell Lines
2
Hepatocyte Differentiation Protocol
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When the cells attained a confluence of 40%, mTeSR™1 was replaced with RPMI 1640 (Gibco), containing 2% B27 minus insulin (Gibco), 100 ng/mL activin A (Peprotech), and 50 ng/mL Wnt 3a (R&D System) for 3 days. The medium was then replaced with IMDM (Procell) with 20% knockout serum replacement (Gibco), 1% GlutaMAX™ supplement (Gibco), 1% dimethyl sulfoxide (DMSO) (Sigma-Aldrich), 1% nonessential amino acids (NEAA) (Gibco), and 0.1 mM β-mercaptoethanol (β-ME) (Sigma-Aldrich) for 4 days. Next, the cultures were placed in IMDM containing 1% GlutaMAX™ supplement, 20 ng/mL oncostatin M (OSM) (Peprotech), 5 ng/mL basic fibroblast growth factor (bFGF) (Peprotech), 0.5 μM dexamethasone (Dex) (Sigma-Aldrich), and 1% insulin-transferrin-selenium (ITS) (Sigma-Aldrich) for 5 days. Finally, the differentiated cells were cultured in hepatocyte culture medium (HCM) (Lonza) supplemented with 10 ng/mL hepatocyte growth factor (HGF) (Peprotech), 0.5 μM Dex, 10 μM lithocholine acid (LCA) (Sigma-Aldrich), 10 μM vitamin K2 (Sigma-Aldrich), and 1% ITS for continuous cultivation.
3
Culturing Human Megakaryocyte Cell Line
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M-07e is a human megakaryocyte cell line, purchased from Cellcook Biotech (Guangzhou, China), which is a suspension cell. These cells were cultured in Iscove’s Modified Dulbecco’s Medium (IMDM) (Procell,China) supplemented with 10% fetal calf serum (FBS) (CellMax, Australia) and 10 ng/ml Granulocyte-macrophage colony stimulating factor (GM-CSF) (Peprotech,USA) in a humidified 5% CO2 incubator at 37 °C.
4
ASP Cytotoxicity on Human Chronic Myeloid Leukemia
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Human chronic myeloid leukemia cells K562 was purchased from Procell Life Science & Technology Co., Ltd. (cat. no. CL-0130, China) and authenticated by STR profiling. Cells were cultured in IMDM (Procell, China) containing 10 % fetal bovine serum (FBS; cat. no. 164210–50, Procell, China) and 1 % Penicillin-Streptomycin Solution (cat. no. PB180120, Procell, China) with an incubator at 37 °C with 5 % CO2.
ASP was obtained from Solarbio (cat. no. SA9770, China). K562 cells were treated with 0.1 % DMSO as the control and 0, 200, 400, 800, 1600 μg/mL ASP for 24 h, separately, and the IC50 of ASP was detected. Subsequent experiments were performed using 1/2 IC50, IC50, and 2 IC50 ASP to test the effects of ASP on K562 cells.
ASP was obtained from Solarbio (cat. no. SA9770, China). K562 cells were treated with 0.1 % DMSO as the control and 0, 200, 400, 800, 1600 μg/mL ASP for 24 h, separately, and the IC50 of ASP was detected. Subsequent experiments were performed using 1/2 IC50, IC50, and 2 IC50 ASP to test the effects of ASP on K562 cells.
5
Knockdown and Overexpression of SH3TC2 and YTHDF1 in CRC Cells
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Two human CRC cell lines, HCT116 and SW480, were obtained from Procell (Wuhan, China) and the National Infrastructure of Cell Line Resource (NICR), respectively. HCT116 and SW480 cells were cultured in McCoy 5A (Procell, China) and IMDM (Procell, China) medium containing 10% FBS, respectively. Cells were maintained in a wet incubator with 5% CO2 at 37°C. The recombinant lentivirus used for SH3TC2 knockdown (sh-SH3TC2#1, #2) and the corresponding control lentivirus (sh-NC) were purchased from GeneChem (Shanghai, China). The overexpression plasmids for wide-type (OE-YTHDF1-WT) and mutant (OE-YTHDF1-Mut) YTHDF1 were also obtained from GeneChem (Shanghai, China). The reagents HiTransG A (GeneChem, China) and Lipo3000 (Thermo, USA) were utilized to transfect lentiviruses and plasmids into CRC cells, respectively.
6
Gastric Cell Line Culture Protocol
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The gastric epithelial cell line GES-1 and GC cell lines (AGS, BGC-823, HGC-27, MKN-28 and KATO III) were provided by Procell Life Science & Technology Co., Ltd. (Wuhan, China). AGS cells were cultured in Ham’s F-12 (Procell, CN), KATO III cells were cultured in IMDM (Procell, CN), and other cells were cultured in RPMI-1640 (Procell, CN). All culture media were supplemented with 10% fetal bovine serum and 1% penicillin/streptomycin solution. All cells were cultured at 37 °C with 5% CO2.
7
CEBPB Knockdown in HCT116 CRC Cells
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The human CRC cell line HCT116 was obtained from Procell (Wuhan, China) and the National Infrastructure of Cell Line Resource (NICR), respectively. HCT116 cells were cultured in McCoy 5A (Procell, China) and IMDM (Procell, China) medium containing 10% FBS, respectively. Cells were maintained in a wet incubator with 5% CO2 at 37°C. The recombinant lentivirus used for CEBPB knockdown (sh‐CEBPB#1, #2) and the corresponding control lentivirus (sh‐NC) were purchased from GeneChem (Shanghai, China). The reagent HiTransG A (GeneChem, China) was utilized to transfect lentiviruses into CRC cells.
8
Rapamycin Sensitivity in AML Cells
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Human marrow stromal cells (HS-5) and AML cells (KG-1 and HL-60) were acquired from the American Type Culture Collection (ATCC, Manassas, VA, USA). HS-5 cells were kept in Dulbecco's modified Eagle's medium (DMEM, Procell Life Science and Technology, Wuhan, China) containing 1% penicillin-streptomycin solution (Procell) and 10% fetal bovine serum (FBS, Procell). KG-1 and HL-60 cells were maintained in Iscove's modified Dulbecco's Medium (IMDM, Procell) containing 1% penicillin-streptomycin solution (Procell) and 20% FBS (Procell). All cells were cultured in a humid incubator with 5% CO 2 at 37°C. Rapamycin was diluted in dimethyl sulfoxide (DMSO, Sigma-Aldrich, St. Louis, MO, USA) to concentrations of 20, 40, 80, and 160 nmol/L. For Rapamycin treatment, KG-1 and HL-60 cells were exposed to various doses of Rapamycin (Sigma-Aldrich) for 24 hours. Cells treated with DMSO (Sigma-Aldrich) were used as a control.
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