human osteogenic sarcoma cells|BNCC338485 |BNCC

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human osteogenic sarcoma cells

Culture medium	McCoy's 5a complete medium: 90% McCoy's 5a + 10% FBS
Subculture procedure	Recovery steps: ① The frozen vial is taken out of liquid nitrogen or -80 ℃ refrigerator and put into PE gloves, quickly submersed into a 37 ℃ water bath pan, shaken The frozen vial to accelerate dissolution, and it is advisable to dissolve all within 1min; (2) Add the dissolved cell solution into a centrifuge tube containing 9ml of complete culture medium in an ultra-clean table, centrifuge at 1000-1200rpm for 5min, discard the supernatant, resuspend the cells with 1-2ml of complete media. (3) The cell suspension was added to T25 flask containing 5-6mL complete medium and cultured in an incubator. cell subculture: ① remove the medium, rinse twice with PBS, and add 1-2mL pancreatin (0.25% Trypsin + 0.02% EDTA); (2) Observe the digestion situation under the microscope. When the cell edge shrinks and the adherent is loose (a pipette can be used to suck up some pancreatin and gently blow somewhere in the cell layer, and the cell layer can be seen to detach with naked eyes, I .e. digestion is completed, otherwise digestion is continued). directly suck out pancreatin, add 5-6mL of complete medium, gently blow the cell layer off. (3) dispense the cell suspension into a fresh T25 flask as a ratio of 1:2, add appropriate complete culture medium, mix the cell suspension evenly, and culture in an incubator.; ④ Pay attention to the change of pH of medium and cell density, renew the medium regularly (2-3 times a week), and repeat the subculture or cryopreservation when the cell density reaches 80%-90%.
Growth conditions	37 ℃;5% CO2 + 95% air;
Growth characteristics	Adherent growth
Storage conditions	Liquid nitrogen
Safety level	1
morphology	Epithelial cell-like, short spindle
Separation substrate	Osteosarcoma, bone.
application	This cell line is suitable as a transfection host.
Sharing mode	Public welfare sharing

1. cell name: Saos-2

2. No. & nbsp;:BNCC 338485

3. growth properties: ■ adherence □ suspension □ semi-suspension and semi-adherence

4. growth conditions:

culture medium	85% McCOY ' S 5A(SIGMA, article number M4892, add NaHCO3 2.2g/L)
serum	10% FBS
temperature	37 ℃
atmosphere	5% CO2,95% AIR
frozen storage conditions	culture medium 50%, serum 40%, DMSO 10%

5. composition:

composition	specifications
frozen via	250 ul/1
handling procedure	1 copy

Attentions and handling procedures

1 upon receipt, please recover the cells immediately if dry ice is exhausted, and do not cryopreserve the cells again; If there is still dry ice, please immediately place the frozen vials into liquid nitrogen, and store the cells according to the specified conditions. Do not put the cells at high temperature.

2 Please recover the cells within 4 weeks after receipt, and check the cell viability. The compliant is not accepted if it is overdue reported.

3 It is not necessary to centrifuge the frozen cell, because the cryopreservation reagent proportion is optimized. After thawing, directly transfer 250ul of cell suspension into the culture vessel containing 8-10 ml of complete medium.

Recovery and subculture handling procedures of adherent cells ( under strict aseptic conditions )

1 Remove the medium in the original culture flask, rinse twice with PBS, and add 1~2 ml of 0.25% EDTA trypsin for digestion (usually in 1~2min).

2 Observe the digestion under the microscope. When the cell edge shrinks and adherent is loose, remove the trypsin, add 6~8ml complete medium, aspirate the cell layer off.

3 Transfer part of the cell suspension to a fresh culture vessel / flask, add appropriate complete medium, and culture it in the incubator.

4 Pay attention to the change of pH value of medium and cell density, renew the medium regularly, and repeat the subculture or cryopreservation when the cell density reaches 70%-80%.

Recovery and subculture handling procedures of suspension cells ( under strict aseptic conditions )

1 The suspension cells is normally handled by dividing the cell solution, subculture in separate flasks, that is, transfer half of the cell suspension to a fresh culture vessel, add appropriate complete medium and cultivate in incubator; It can also be divided and subcultured in several flasks according to cell density.

2 Pay attention to the change of pH value of medium and cell density, renew the medium regularly, and repeat the subculture or cryopreservation when the cell density reaches 70%-80%.

Handling procedures of semi-adherent and semi-suspended cells ( under strict aseptic conditions )

1 If there are many suspended cells with good refractive index, they can be collected by centrifugation and continue to be cultured.

2 If there are few suspended cells, it may not be collected. And subculture according to the handling procedures of adherent cells

3 If there are many suspended cells, collect them by centrifugation. The adherent cells in the flask are operated according to the handling instructions of adherent cell for digestion, termination of digestion, aspiration. Suspend them with the previously collected suspension cells and dispense into separate culture vessels.