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MB Sample ID: SA085208
Local Sample ID: | K5 |
Subject ID: | SU001277 |
Subject Type: | Cultured cells |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606 |
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Treatment:
Treatment ID: | TR001292 |
Treatment Summary: | Cells were split when they reached a confluency of 80 %. Passage 3 or 4 were used. Cells were plated in 20 Petri dishes (10 cm2, 1 x 106 cells/ per dish) and cultivated for 4 days in 10 mL of the above mentioned medium for adherence. 10 dishes were incubated with 1.95 µM betulin (10 µL of the 1.95 mM stock solution in DMSO) for 8 hrs prior to removal of the medium and the remaining ones were used as control. Control samples were treated with 10 µL DMSO. The concentration of 1.95 µM of betulin was used, as this concentration has shown effects in our previous studies on the molecular wound healing effect [1]. On day 4, cells were incubated with 3 ml of trypsin (0.05 %) at 37°C. After 5 min cells were washed by adding 7 mL medium. The suspension was transferred into 15 mL falcon tubes and centrifuged for 5 min at 4°C and 1.200 rpm, respectively. The supernatant was withdrawn and the remaining cell pellets were washed and then frozen at -20°C in the falcon tubes until extraction. |
Treatment Protocol Filename: | ccalcas_Extraction_and_treatment.pdf |