||Mixed, Adherent And Suspension
||4 Year /
||was established in 1970 from a metastatic bone tumor.
||This cell line is suitable as a transfection host.
||SH-SY5Y cells have a reported saturation density greater than 1 X 10e6 cells/cm2. They are reported to exhibit moderate levels of dopamine beta hydroxylase activity. [ PubMed: 29704]
||1:1 mixture of Dulbecco's modified Eagle's medium and F12 Medium containing 1% non-essential amino acids, 2 mM L-glutamine, 1 mM sodium pyruvate and fetal bovine serum to a final concentration of 10%.
||These cells grow as a mixture of floating and adherent cells. The cells grow as clusters of neuroblastic cells with multiple, short, fine cell processes (neurites). Cells will aggregate, form clumps and float.
Remove the medium with the floating cells, and recover the cells by centrifugation. Rinse the adherent cells with PBS without calcium and magnesium, add an additional 1 to 2 mL of trypsin solution, and let the culture sit at room temperature (or at 37°C) until the cells detach.
Add fresh medium, aspirate, combine with the floating cells recovered above and dispense into new flasks.
NOTE: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 12 in Culture of Animal Cells, a manual of Basic Technique by R. Ian Freshney, 6th edition, published by Alan R. Liss, N.Y., 2010.
|Subculturing Medium Renewal
||Every 4 to 7 days
|Subculturing Subcultivation Ratio
||1:20 to 1:50 is recommended
||Atmosphere: air, 95%; carbon dioxide (CO2), 5%
||95% FBS + 5% DMSO (Dimethyl sulfoxide)
|Thawing Frozen Cells
||SAFETY PRECAUTION: Is highly recommend that protective gloves and clothing always be used and a full face mask always be worn when handling frozen vials. It is important to note that
some vials leak when submersed in liquid nitrogen and will slowly fill with liquid nitrogen. Upon thawing, the conversion of the liquid nitrogen back to its gas phase may result in the vessel exploding or blowing off its cap with dangerous force creating flying debris.
1. Thaw the vial by gentle agitation in a 37°C water bath. To reduce the possibility of contamination, keep the Oring and cap out of the water. Thawing should be rapid (approximately 2 minutes).
2. Remove the vial from the water bath as soon as the contents are thawed, and decontaminate by dipping in or spraying with 70% ethanol. All of the operations from this point on should be carried out under strict aseptic conditions.
3. For cells that are sensitive to DMSO is recommended that the cryoprotective agent be removed immediately. Transfer the vial contents to a centrifuge tube containing 9.0 mL complete culture medium and spin at approximately 125 x g for 5 to 7 minutes.
4.Discard the supernatant and Resuspend cell pellet with the recommended complete medium (see the specific batch information for the culture recommended dilution ratio).
5. Incubate the culture in a appropriate atmosphere and temperature (see "Culture Conditions" for this cell line).
NOTE: It is important to avoid excessive alkalinity of the medium during recovery of the cells. It is suggested that, prior to the addition of the vial contents, the culture vessel containing the growth medium be placed into the incubator for at least 15 minutes to allow the medium to reach its normal pH (7.0 to 7.6).
||22554: Ross RA, et al. Coordinate morphological and biochemical interconversion of human neuroblastoma cells. J. Natl. Cancer Inst. 71: 741-749, 1983. PubMed: 6137586
23032: Biedler JL, et al. Multiple neurotransmitter synthesis by human neuroblastoma cel
||CAROLINA ALVARES DA CUNHA - Universidade Federal do Rio de Janeiro
Flávio Pereira Kapczinski - Universidade Federal do Rio Grande do Sul