Materials and Reagents

1. 12 well cell culture plate (Greiner Bio One International GmbH, catalog number: 665180 )
   
2. 96 well tissue culture test plate (TPP Techno Plastic Products AG, catalog number: 92696 )
   
3. 1.5 ml tubes (Carl Roth GmbH + Co., catalog number: 7080.1 )
   
4. ThinCerts^TM cell culture inserts with a pore diameter of 8 µm (Greiner Bio-One GmbH, catalog number: 665638 )
   
5. Cover slips for haemocytometer (Carl Roth GmbH + Co., catalog number: L189.1 )
   
6. Filter tips (10 µl, 20 µl, 100 µl, 200 µl and 1,000 µl) (Carl Roth GmbH + Co., catalog number: 771288 , 774288 , 772288 , 739288 and 740288 )
   
7. Tumor cells (the lung cancer cell lines H1975 and 2106T)
   
8. Cell culture medium (cell line specific), with and without fetal bovine serum (FBS)
   
9. Heat-inactivated FBS (E.U.-approved, South America Origin) (Thermo Fischer Scientific, catalog number: 10500-064 )
   
10. 1x Dulbecco’s phosphate-buffered saline (DPBS, no calcium, no magnesium) (Thermo Fischer Scientific, Gibco^TM, catalog number: 14190-094 )
    
11. StemPro^® Accutase^® cell dissociation reagent (Thermo Fischer Scientific, Gibco^TM, catalog number: A11105-01 )
    
12. Mitomycin C (AppliChem GmbH, catalog number: A2190,0002 )
    
13. Trypan blue solution (Sigma-Aldrich, catalog number: T8154 )
    
14. 10x cell lysis buffer (Cell Signaling Technology, catalog number: 9803S )
    
15. Cytotoxicity detection kit (LDH) (Roche Diagnostics, catalog number: 11644793001 )
    

Equipment

1. CO₂ cell incubator (Panasonic Corporation, Sanyo, model: SANYO-InCu saFe^® )
   
2. Neubauer cell counting chamber (VWR International, catalog number: BRND717810 )
   
3. Microcentrifuge (Eppendorf AG, model: 5417R )
   
4. Vortex mixer (VWR International, catalog number: 444-1372 )
   
5. Versatile microplate absorbance reader (Tecan Trading AG, model: sunrise^TM )
   
6. Gilson’s Pipetman classic pipettes ( P10 , P20 , P100 , P200 , P1000 )
   

Procedure