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A549-DDP

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Cat. No.: CL-0519 Publications(73)
A549-DDP - 1
  • A549-DDPA549-DDP - 1
  • A549-DDPA549-DDP - 2
  • A549-DDPA549-DDP - 3
  • +2

CL-0519

$ 500.00

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1×10^6Cells/Vial×2Vials

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Product Introduction

General information
Cell Name A549-DDP
Organism Human
Growth Properties Adherent
Morphology Epithelial-like
Disease lung cancer cells
Tissue Lung
Age 58Y
Sex Male
Instructions 1. Check all containers for leakage or breakage.
2. Remove the frozen cells from the dry ice packaging and immediately transfer them to liquid nitrogen (liquid or vapor phase) for long-term cryopreservation.
3. Drug-resistant cell lines display slow proliferation. When subcultured at a 1:2 split ratio, cells are ready for the next passage in approximately one week, and they show an elongated spindle morphology. If the cells proliferate extremely slowly, perform expansion culture following the procedure below: Culture cells continuously in medium with a low drug concentration for two weeks. Carry out subcultivation whenever required based on cell confluency during this period. After cell growth status recovers, gradually increase the drug concentration in a stepwise gradient. Maintain cells at each concentration for one full passage—culture until reaching full confluency and split cells before switching to the next higher concentration. Once stable cell growth and morphology are confirmed, maintain continuous culture in drug-supplemented medium.
Complete Medium Ham's F-12K [PM150910]+10% Nutrient+1 μg/mL Supplement1+1% Supplement2
Incubation Atmosphere Air, 95%; CO₂, 5%
Temperature 37℃
Subcultivation Ratio 1:2-1:3
Medium Renewal 2 to 3 times per week
Dissociation Duration 2-3 min
Subculturing Procedure 1. Remove the culture medium from the T25 cell culture flask.
2. Add approximately 2 mL of PBS. Gently tilt the flask side to side until the PBS covers the entire bottom, then aspirate and discard the PBS.
3. Add 1 mL of 0.25% trypsin solution (containing EDTA). Gently tilt the flask side to side until the trypsin solution covers the entire bottom of the flask.
4. Incubate the cells at 37°C. Observe the cells under an inverted microscope and terminate digestion once the cells round up and detach. To avoid clumping, do not agitate the cells by tapping or shaking the flask during detachment.
5. Add 3 mL of complete culture medium to terminate digestion and disperse into a single cell suspension.
6. Collect the cell suspension and centrifuge at 1200 rpm (approximately 250 ×g) for 3 minutes. Carefully aspirate and discard the supernatant.
7. Add fresh complete culture medium, pipette gently several times to resuspend the cells, and seed them at the appropriate ratio into a new culture flask. Loosen the cap or use a vented cap for incubation.
Freeze Medium General Freezing Medium [PB180436]
Storage Conditions For long-term cryopreservation, cryovials should be stored in liquid nitrogen at −150°C to −196°C. Storage at −80°C is restricted to short-term interim use only.
Background The A549-DDP cell line is a cisplatin (DDP)-resistant subline derived from parental A549 cells using the dose-escalation method.
Biosafety Level BSL-1

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