changes in protein profiles in bortezomib applied multiple myeloma ...

3.1. Materials
3.1.1. Medias
CHAPTER 3
MATERIALS AND METHODS
Medias were listed in Appendix A.
3.1.2. Chemicals, Reagents and Solutions
Reagents and solutions were presented in Appendix B.
3.2. Methods
3.2.1. Cell Lines and Culture Conditions
U-266 human MM cells were kindly obtained from the German Collection of
Microorganisms and Cell Cultures (Germany). The cells were grown and maintained in
RPMI-1640 growth medium containing 10% fetal bovine serum (FBS) and 1%
penicillin-streptomycin (Invitrogen) at 37°C in 5% CO2. Medium was refreshed every 3
days. In order to passage the cells, whole cell suspension was taken from tissue culture
flask (75cm 2 ) into a sterile falcon tube (50ml) and then centrifuged at 1000 rpm (~800g)
for 10 minutes (min) at room temperature. After centrifugation, the supernatant was
removed and the pellet was washed with 3-4 mL Phosphate Buffered Saline (PBS) and
centrifugation process was repeated. In the following step, supernatant was removed
from the tube and the pellet was resuspended with 15 mL of RPMI-1640 (10% FBS and
1% Penicillin-Streptomycin). Then, it was transferred into a sterile 75cm 2 filtered tissue
culture flask and incubated in CO2 incubator.
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