human small cell lung cancer cells 

DMS153

BNCC number: 102040

Growth properties: Suspension growth

culture conditions: 37 ℃,5% CO₂

complete medium: 90% EMEM + 20% FBS

frozen storage conditions: 50% EMEM + 40% FBS + 10% DMSO

Receiving notice:  if any abnormality is found on the day of receipt, please contact the customer service within 24 hours. If it is overdue, it is deemed that the cells are well. For the frozen cells,  it shall be stored in the refrigerator at -80 ℃ upon arrival. If they are not used for a long time, they shall be transferred to liquid nitrogen for storage overnight.  Recovered cells in T25 culture flask, upon receipt, put the culture flask in the incubator for 2-3 hours, and then carry out handling procedure. Under sterile conditions, centrifuge the cell solution at 1000-1200rpm/min for 5-10 minutes, resuspend the cells, transfer the suspended cells in the T25 flask, add 10ml of complete medium, then put it in incubator. Please operate in strict accordance with this instruction, otherwise the replacement of cells are not be available in case of loss of cell viability.

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, shaken the frozen vial to accelerate dissolution, and it is advisable to dissolve all within 1min; 
(2)    Put 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 6-8ml complete medium and cultured in an incubator. 

cell subculture:

① Centrifugation method: Collect the cells, centrifuge at 1000rpm for 5 minutes, discard the supernatant, add 1-2mL of culture medium and blow evenly, divide the cell suspension into a new medium containing 8ml of medium at a ratio of 1:2. in a T25 bottle.

②Half-volume medium exchange method: After the culture flask is left standing for 5-10 minutes, half of the supernatant medium is gently sucked away, and the remaining medium with cell sediment is resuspended and evenly mixed, and the cell suspension is divided at a ratio of 1:2. into a new T25 flask containing 8 ml of medium.

③ Pay attention to the pH value of the medium and the cell density, and change the medium regularly (2-3 times a week). When the cell density reaches more than 2×106 cells/ml, repeat the passage or cryopreservation.

Notes:
(1)    The cells are density dependent. Initiate the subcultivation in T25 flask when the cell density reaches 80%; A subcultivation ratio of 1:2 is recommended. 
(2)    If the cells in sealed culture bottle, put it into the incubator for cultivation after handling, with the cap unscrewed.

Recovery record:  According to the recovery instructions, the results of the cell recovery are reported as follows:

item	quality standard	Recovery record
Viability:	suspension growth rate ≥ 80.0% in 118 hours	suspension growth rate reaches  30.0% in 18 hours,  and 80.0% in 126 hours
cell morphology:	suspension	Suspended, round, multiple clusters, a small amount of loose adherence
attached figure:
Conclusion:	good viability, no abnormal cell morphology, qualified