SH-SY5Y and U251 cells (ATCC, Manassas, VA, US) were maintained on tissue culture dishes coated with type I collagens (Nitta Gelatin, Tokyo, Japan) and grown in low glucose DMEM (Wako, Tokyo, Japan) added with 10% fetal bovine serum (Thermo Fisher Scientific, Tokyo, Japan) and appropriate antibiotics (Wako, Tokyo, Japan). For transfection, SH-SY5Y and U251 cells were inoculated into 24-well tissue culture plates and then transfected with NC, miR-125a precursor, or ET1 siRNA by Fugene 6 HD (Promega, Madison, WI). At 48 h after the transfection, transfected cells were harvested to assay the expression levels of ET1. Data were averaged from 3 repeated experiments.
Sh sy5y
Manufactured by Promega
SH-SY5Y is a human-derived neuroblastoma cell line. It is commonly used as a model for neurological research and studies.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Dmem low glucose, by Fujifilm (1 mentions)
Fugene 6 hd, by Promega (1 mentions)
Ham s f 12, by Merck Group (1 mentions)
Glutamax, by Thermo Fisher Scientific (1 mentions)
Minimum essential medium (mem), by Merck Group (1 mentions)
Antibiotic antimycotic, by Thermo Fisher Scientific (1 mentions)
Sh sy5y, by European Collection of Cell Cultures (1 mentions)
Methyl thiazolyl tetrazolium (mtt), by Promega (1 mentions)
2 protocols using sh sy5y
1
SH-SY5Y and U251 Cells Transfection
2
Viability Assay for Human Neuroblastoma Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Human neuroblastoma cells (SH-SY5Y)
were obtained from the European Collection of Cell Cultures (ECACC,
Sigma-Aldrich, Dorset, U.K.) and grown in a 1:1 mixture of minimal
essential medium (MEM) (Sigma-Aldrich) and nutrient mixture Ham’s
F-12 (Sigma-Aldrich) supplemented with 15% FBS, 1% non-essential amino
acids, 2 mM GlutaMAX, and 1% antibiotic-antimycotic (all from Thermo
Fisher Scientific, Epsom, U.K.). The vitality of SH-SY5Y cells after
treatment with aSyn was determined using an MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium
bromide] kit from Promega (Madison, WI). Live cells with active metabolism
convert MTT to formazan. Cells (3 × 104) were seeded
into each well of 96-well culture plates overnight followed by incubation
of aSyn for 24 h. MTT was added to cells for 4 h, and the absorption
was measured with a microplate reader at 590 nm (Envision, PerkinElmer).
The absorbance is proportional to the number of viable cells; therefore
>100% vitality may be due to an increase in the number of cell
compared
to the control as these are nondifferentiated neuroblastoma cells.
Each experiment was performed three times using duplicates for aSyn
samples from each purification batch.
were obtained from the European Collection of Cell Cultures (ECACC,
Sigma-Aldrich, Dorset, U.K.) and grown in a 1:1 mixture of minimal
essential medium (MEM) (Sigma-Aldrich) and nutrient mixture Ham’s
F-12 (Sigma-Aldrich) supplemented with 15% FBS, 1% non-essential amino
acids, 2 mM GlutaMAX, and 1% antibiotic-antimycotic (all from Thermo
Fisher Scientific, Epsom, U.K.). The vitality of SH-SY5Y cells after
treatment with aSyn was determined using an MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium
bromide] kit from Promega (Madison, WI). Live cells with active metabolism
convert MTT to formazan. Cells (3 × 104) were seeded
into each well of 96-well culture plates overnight followed by incubation
of aSyn for 24 h. MTT was added to cells for 4 h, and the absorption
was measured with a microplate reader at 590 nm (Envision, PerkinElmer).
The absorbance is proportional to the number of viable cells; therefore
>100% vitality may be due to an increase in the number of cell
compared
to the control as these are nondifferentiated neuroblastoma cells.
Each experiment was performed three times using duplicates for aSyn
samples from each purification batch.
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