Viruses |
MAP-test: Negative (Sendai, Ektromelie, Polyoma, K-Virus, Kilham, Reo 3, PVM, LCM, M.pulmonis, MVM, Theilers GD VII, Toolans H-1, MHV, LDV, RCV/SDA, M-Adenovirus, B.piliformis). |
Subculturing |
Remove medium and rinse the adherent cells using PBS without calcium and magnesium (3-5 ml PBS for T25, 5-10 ml for T75 cell culture flasks). Add Accutase (1-2 ml per T25, 2.5 ml per T75 cell culture flask), the cell sheet must be covered completely. Incubate at ambient temperature for 8-10 minutes. Carefully resuspend the cells with medium (10 ml), centrifuge for 3 min at 300 g, resuspend cells in fresh medium and dispense into new flasks which contain fresh medium. Using collagen-coated flasks increases the number of adherent cells. |