K562 cells were cultured in RPMI-1640 medium (Sigma‒Aldrich, United States) containing 10% fetal bovine serum (FBS, Sigma‒Aldrich, United States) and 1% penicillin and streptomycin (Sigma‒Aldrich, United States). K562 cells were differentiated by erythroid induction at an initial concentration of 1 × 105/mL cells. A final concentration of 40 μM hemin (Solarbio, China) and 0.1 ng/mL cytarabine (Solarbio, China) were added to the medium to induce erythroid differentiation, and the cells were placed at 37°C under hypoxic (5% CO2, 5% O2), CoCl (200 μM) and normoxic (5% CO2, 20% O2) conditions for 5 days.
Cytarabine
Manufactured by Solarbio
Sourced in China
Cytarabine is a synthetic nucleoside analog used in laboratory settings for research purposes. It functions as an antimetabolite, inhibiting DNA synthesis and cellular division. Cytarabine is a widely used tool in cell biology and cancer research.
Automatically generated - may contain errors
Lab products found in correlation
Hemin, by Solarbio (1 mentions)
Penicillin and streptomycin, by Merck Group (1 mentions)
Fetal bovine serum (fbs), by Merck Group (1 mentions)
Rpmi 1640 medium, by Merck Group (1 mentions)
Trypsin, by Avantor (1 mentions)
Collagenase 1, by Solarbio (1 mentions)
D hank s solution, by Merck Group (1 mentions)
Dmem medium, by Merck Group (1 mentions)
Cytarabine ara c, by Merck Group (1 mentions)
L glutamine, by Solarbio (1 mentions)
Trypan blue, by Solarbio (1 mentions)
Rat tail collagen type 1, by Thermo Fisher Scientific (1 mentions)
L 15 medium, by Thermo Fisher Scientific (1 mentions)
Daunorubicin, by Solarbio (1 mentions)
5 protocols using cytarabine
1
K562 Cell Erythroid Differentiation
2
Neuronal Cell Viability Assay
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N-methyl-D-aspartic acid (NMDA, Shanghai Yuanye Biology company); DMEM medium (Sigma-Aldrich company); trypsin (Amresco company); D-Hank’s solution (Sigma-Aldrich company); cytarabine (Ara-C, Sigma-Aldrich company); CCK-8 Kit (Shanghai Bogu Biotechnology Co., Ltd.); malondialdehyde kit (Nanjing Jiancheng Bioengineering Research Institute); superoxide dismutase kit (SOD, Nanjing Jiancheng Biology); GSH-PH Kit (Nanjing Jiancheng Bioengineering Institute); DMEM/F-12 medium (Hyclone company); PBS solution (1x, Solarbio company); trypan blue (Solarbio company); collagenase I (Solarbio company); cytarabine (Solarbio company); L-glutamine (Solarbio company).
3
Culturing Primary DRG Neurons with Breast Cancer Cells
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Primary cultures of DRG were established according to a published protocol [21 (link)]. Two days old Sprague Dawley rats were obtained from the Experimental Animal Center of Dalian medical university (permit number SCXK 2008-0002). Briefly, DRG was dissected from newborn Sprague Dawley (2 days old) and pooled in an L-15 medium (Gibco) on ice. The DRG was then transferred to the lower chamber of the Transwell which was covered with rat tail collagen Type I (Invitrogen, USA). After 24 h, the DRG neurons were purified with 5 μg/ml cytarabine (Beijing Solarbio Science & Technology Co., Ltd., Beijing, China). MDA-MB-231 cells were cultured in the upper chambers of the 6-well plates at a density of 1 × 104 cells/cm2. The control group contained only the DRG (referred to as the DRG group). The cells were cultured at 37°C in a 5% CO2 incubator for 7 days and the medium was changed every 2 days. Using NE, NGF, and β2-AR/TrkA blocker (Shandong Topscience, Rizhao, China) pretreatment, the axon growth of each group of DRG neurons was detected by immunofluorescence staining.
4
Isolation and Culture of Mature Neurons
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After removing the brain from fetal rats, the cerebral cortex tissue was carefully separated and cut. Ethylene Diamine Tetraacetic Acid (EDTA)-0.05% trypsin (SolarBio, Beijing) was added for digestion at 37°C for 30 min. Digestion was stopped by adding 20% Fetal Bovine Serum-Dulbecco’s Modified Eagle Medium (FBS-DMEM) complete medium (Procell, Wuhan). The cells were passed through a 70 μm cell sieve, centrifuged at 1500 rpm for 5 min and resuspended in complete medium. The cells were placed in coated bottles and the medium was changed every 3 days. On day 7, 5 μg/ml cytarabine (Solar Bio, Beijing) was added for 72 h to inhibit cell proliferation other than neurons. Mature neurons were then utilized for in vitro trials.
5
Cell Proliferation and Viability Assay
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Cell proliferation was determined with the Cell Counting Kit-8 (CCK8, Dojin Laboratories, Kumamoto, Japan) assay. Briefly, 4 × 10E+4 cells were seeded into each well of 96-well plates. 2, 4 or 7 d later 10 μl of the kit reagent was added to each well and 2 h later all plates were scanned by a microplate reader at 450 nm. CCK8 was also used to determine cell viability after drug exposures including daunorubicin, dexamethasone, methotrexate, cytarabine and imatinib (Solarbio, Beijing, China). Cells were seeded and 72 h later 10 μl of the kit reagent was added to each well and 2 h later plates were scanned by a microplate reader at 450 nm. Cell viability was assessed based on the value of fluorescent signal of live cells with no drug treatment. Experiments were performed in triplicate for 3 times independently.
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