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【求助】胶束如何做MTT
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The cytotoxicity of the L121 micelles (uncrosslinked), L121-CHO micelles, and crosslinked L121 micelles was evaluated in vitro using the MTT assay. The assay is based on the mitochondrial dehydrogenase cell activity as an indicator of cell viability. Briefly, MTT [3-(4,5- dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] was dissolved in phosphate buffered saline (PBS) to a concentration of 5mg/ml as a stock MTT solution and filtered for sterilization. Hs68 cells (human foreskin fibroblasts) were seeded in 24-well plates at 5104 cells/well and were allowed to adhere overnight. The growth medium was replaced with a fresh one containing 0–0.2wt% test micelles. The cells were then incubated for 24 h and washed twice in 1ml PBS. Subsequently, the cells were incubated in a growth medium that contained 1mg/ml MTT reagent for an additional 4 h at 37 1C and 500 ml of dimethyl sulfoxide was added to each well to ensure solubilization of formazan crystals. Finally, the optical density readings were taken using a multiwell scanning spectrophotometer (MRX Microplate Reader, Dynatech Laboratories Inc., Chantilly, VA, USA) at a wavelength of 570 nm.  |
2楼2009-05-27 11:18:52
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