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hanyan2004銀蟲(chóng) (初入文壇)
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胰島素增敏活性篩選實(shí)驗(yàn)操作翻譯
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胰島素增敏活性篩選實(shí)驗(yàn) 用含10% FBS的高糖有酚紅DMEM 培養(yǎng)基在六孔板中培養(yǎng)3T3-L1細(xì)胞，至匯合度達(dá)90%進(jìn)行誘導(dǎo)分化，用含10% FBS，Ins, Dex, IBMX 的高糖有酚紅DMEM 培養(yǎng)基誘導(dǎo)，四天后重新消化接種于24孔板，用含10% FBS的高糖有酚紅DMEM 培養(yǎng)基培養(yǎng)兩天。然后再用含10% FBS和較低濃度Ins較高濃度 Dex的高糖有酚紅DMEM 培養(yǎng)基進(jìn)行誘導(dǎo)胰島素抵抗，四天后換用含待測(cè)樣品的高糖無(wú)酚紅DMEM培養(yǎng)基，同時(shí)設(shè)立空白對(duì)照(誘導(dǎo)分化后的正常細(xì)胞)和陰性對(duì)照(誘導(dǎo)分化后的胰島素抗性細(xì)胞)不加樣品。繼續(xù)培養(yǎng)50h后取10ul 培養(yǎng)上清, 轉(zhuǎn)到酶標(biāo)板中，采用葡萄糖檢測(cè)試劑盒檢測(cè)葡萄糖水平，490nM測(cè)定OD值。實(shí)驗(yàn)時(shí)，每個(gè)樣品設(shè)雙復(fù)孔，重復(fù)兩次。根據(jù)OD值計(jì)算增敏率，增敏率=[1-(OD樣品-OD空白)/(OD陰性-OD空白)] ×100%，相對(duì)增敏率=樣品增敏率/羅格列酮增敏率。 |
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jingjing1005
木蟲(chóng) (小有名氣)
- 翻譯EPI: 27
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wypward(金幣+1):歡迎新蟲(chóng)，辛苦啦 2010-06-01 10:51:07
hanyan2004(金幣+50, 翻譯EPI+1): 2010-06-01 23:17:06
hanyan2004(金幣+1): 2010-06-02 12:10:53
hanyan2004(金幣+1):翻譯得很好，謝謝你哈 2010-06-02 21:37:14
wypward(金幣+1):歡迎新蟲(chóng)，辛苦啦 2010-06-01 10:51:07
hanyan2004(金幣+50, 翻譯EPI+1): 2010-06-01 23:17:06
hanyan2004(金幣+1): 2010-06-02 12:10:53
hanyan2004(金幣+1):翻譯得很好，謝謝你哈 2010-06-02 21:37:14
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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. |
2樓2010-06-01 10:22:34
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