Human gastric cancer cells|BNCC340918 |BNCC

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Human gastric cancer cells

Culture medium	IMDM complete medium: 90% IMDM + 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. ③The cell suspension was then added to the T25 flask containing 6-8mL of complete medium and placed in an incubator for culture. cell subculture: ① transfer the cell suspension solution into a centrifuge tube, centrifuge at 1000rpm for 5min, and collect cells; (2) rinse T25 flask twice with PBS, add 1-2mL pancreatin (0.25% Trypsin + 0.02% EDTA); Observe the digestion situation under the microscope. When the cell edge shrinks and the adherent is loose (a pasteur pipette can be used to suck up some pancreatin, and gently blow somewhere in the cell layer, , the cell layer can be seen to detach with the naked eye, that is, digestion is completed, otherwise digestion continues), directly suck out pancreatin, add 5-6mL of complete medium, gently blow the cell layer off. (3) The cells obtained from the above two steps are evenly mixed and then dispensed into fresh T25 flasks as a ratio of 1:2. add appropriate complete 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	Culture temperature 37 ℃; Atmosphere 5% CO2 + 95% air;
Growth characteristics	semi-adherent and semi-suspension growth
Storage conditions	Liquid nitrogen
Safety level	1
morphology	Epithelial cell-like, short fusiform, round, irregular margin, clean background, no pigment, no vacuoles
Separation substrate	Stomach
Sharing mode	Public welfare sharing

1. Name:KATO III

2.No.:340918

3. Properties : □ wall □ suspension ■ semi-suspension and semi-wall

4. Growth conditions:

culture medium	90% IMDM
serum	10% FBS(Diagnovum )
temperature	37 ℃
air condition	5% CO₂,95% AIR
growth algebra	P4-5
frozen storage conditions	culture medium 50%, serum 40%, DMSO 10%

5. Composition:

composition	specifications
a bottle of cells	T25
cell culture and operating instructions	1 copy

Receiving notice:

1 Upon arrival, it is suggested to sit the cells in a incubator for about 4 hours, and then renew the media for recovery or subculture according to the cell density.
2 If the adherent cells are received in the form of ( partial) suspension, please centrifuge the suspended cells in time, add 15% serum complete medium to a fresh culture dish / vial and continue to culture for 3 days; At the same time, the remaining adherent cells in the original culture flask were renewed with 15% serum complete medium and cultured for 2-3 days. If the cells do not proliferate after 3 days but continue to detach and die, please contact the technicians.
3 If there are few suspended cells, it may not be collected. And subculture according to the handling procedures of adherent cells

4 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.

Recovery and subculture procedure ( 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 for trypsin digestion (usually in 1~2min)
2 Observe the digestion under the microscope. When the cell edge shrinks and adherent is loose (but not floating), 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%.

Special attention: (if handle in public laboratory or first time to culture the cells, it is recommended to add penicillin streptomycin into the media)
1 Upon the receipt of the cells, renew the solution with fresh 10% FBS medium as soon as possible. It is not recommended to use the medium used for transportation in the original bottle.
2 Please take photos and contact the technicians in time if the culture flask leaks upon the receipt of cells.
3 Any compliant on the cells, please take photos and contctat our technicians.

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