|
|
★
wypward(金币+1):欢迎新虫，辛苦啦 2010-06-01 10:51:07
hanyan2004(金币+50, 翻译EPI+1): 2010-06-01 23:17:06
hanyan2004(金币+1): 2010-06-02 12:10:53
hanyan2004(金币+1):翻译得很好，谢谢你哈 2010-06-02 21:37:14
Screening test of Insulin-sensitizing activity
3T3-L1 cells were cultured in high glucose DMEM culture meduim with phenol red Containing 10% FBS in six-well plates. When confluence reached 90%, cells were induced to differentiate with a high sugar DMEM culture medium with phenol red containing 10% FBS, Ins, Dex, IBMX. After four days, cells were seeded in 24 well plates after re-digestion, cultured in high glucose DMEM culture medium with phenol red containing 10% FBS for two days. and then they were induced for insulin resistance using high glucose DMEM culture medium with phenol red containing 10% FBS, Ins in lower concentration and Dex in higher concentration. Four days later the cells were transferred to the high glucose DMEM medium without phenol red containing sample, while the control test (normal cells after induction of differentiation) and negative control test (insulin resistant cells after induced differentiation without sample) were established. 10ul supernatants were taken to ELISA plate
after 50hs’ continuous culture to test the glucose level using glucose testing kit and OD values in 490nm. During the experiment, a double recovery was set for each sample hole and the same experiment was repeated twice. The increased sensitivity rate was calculated according to OD values, increased sensitivity rate = [1 - (OD sample-OD blank) / (OD negative-OD blank)] × 100%, the relative increased sensitivity rate = increased sensitivity of sample / increased sensitivity of rosiglitazone. |
|
回复此楼