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【公司名稱】廣州健侖生物科技有限公司

【市場(chǎng)部】楊永漢
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【騰訊】
【公司地址】廣州市清華科技園健新基地番禺石樓鎮(zhèn)健啟路63號(hào)二期2幢101-103室

腹腔中分離1、取6周左右的小鼠（或600克左右的豚鼠，或 3公斤左右的家兔），剃去腹部的毛并消毒。腹腔注射1ml（豚鼠 20ml，家兔200ml）無(wú)菌的液體石蠟或巰基乙酸肉湯或4%淀粉肉湯。3～4天以后收集腹腔細(xì)胞。2、如要收集腹腔靜置巨噬細(xì)胞，不注射激物，直接從一步開始。放血處死動(dòng)物，以減少腹腔中的紅細(xì)胞。消毒腹部，沿腹中線注入3～4ml（豚鼠20～40ml，家兔50 ～70ml）冰中預(yù)冷的無(wú)菌PBS-H（含10U/ml肝素和10%小牛血清的 PBS）。輕輕腹部5分鐘。3、以下均無(wú)菌操作。剪開腹壁，用吸管吸出滲出液，再用同樣容量的預(yù)冷PBS-H沖洗腹腔2～3次。合并滲出液于離心管中，4℃離心250×g10分鐘，去上清液。4、用預(yù)冷的RPMI-1640培養(yǎng)液洗滌細(xì)胞3次，每次4℃離心250×g10分鐘，去上清液。用預(yù)冷的適量RPMI-1640培養(yǎng)液懸浮細(xì)胞，臺(tái)盼藍(lán)染色計(jì)數(shù)細(xì)胞并決定細(xì)胞活力。
Isolation in the peritoneal cavity 1. Take about 6 weeks of mice (or about 600 grams of guinea pigs or about 3 kg of rabbits), shave the abdomen and disinfect it. Intraperitoneal injection of 1 ml (guinea pig 20 ml, 200 ml rabbit) sterile liquid paraffin or thioglycollate broth or 4% starch broth. The peritoneal cells were collected 3 to 4 days later. 2. If you want to collect the macrophages in the abdominal cavity without injection, start directly from one step. Blood was sacrificed to reduce red cells in the abdominal cavity. The abdomen was disinfected, and 3 to 4 ml (guinea pig 20 to 40 ml, rabbit 50 to 70 ml) in ice was preincubated in the ice along a midline of the abdomen, pre-cooled in iced PBS-H (PBS containing 10 U/ml heparin and 10% calf serum). Gently abdomen for 5 minutes. 3, the following are aseptic. The abdominal wall was dissected, the exudate was aspirated with a pipette, and the abdominal cavity was washed 2 to 3 times with the same volume of pre-cooled PBS-H. Combine the exudate in a centrifuge tube, centrifuge at 250×g for 10 minutes at 4°C, and remove the supernatant. 4. Wash the cells three times with pre-chilled RPMI-1640 medium, centrifuge at 250 x g for 10 minutes at 4°C each time, and remove the supernatant. Resuspend cells in pre-cooled, appropriate RPMI-1640 medium and trypan blue staining. Count cells and determine cell viability.

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