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廣州健侖生物科技有限公司

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保質期：2年

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【公司名稱】 廣州健侖生物科技有限公司

【 市場部 】       楊永漢
【】 
【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號二期2幢101-103室

家兔肺泡巨噬細胞的分離和純化巨噬 細胞的分離和純化1、取2～3公斤重的家兔，耳緣靜脈注射10ml空 氣處死動物或注射2ml（含130mg）戊巴比妥麻醉動物。仰臥固定 ，消毒，剪開胸壁。以下過程注意無菌操作。小心從環(huán)狀軟骨下 方剪下氣管，分離心臟和血管，取出肺，剪去脂肪和結締組織。 用無菌紗布小心擦凈肺表面，稱重。2、分離肺泡巨噬細胞：用夾 子夾住氣管，使肺懸空。向氣管中注射40ml無菌的冷生理鹽水， 讓生理鹽水進入全部肺泡。用鑷子提起氣管，從夾子下面剪斷氣 管，將肺中的液體倒入離心管中。再用同樣方法，用40ml無菌冷 生理鹽水沖洗肺泡，合并浸出液，置4℃。3、分離肺組織中的巨 噬細胞：取出肺泡巨噬細胞后，剪去氣管，將肺組織放在有冷 RPMI-1640培養(yǎng)液的平皿中。平皿置冰上，用吸滿冷RPMI-1640培 養(yǎng)液的20ml注射器接23號針頭，一邊灌注一邊用針頭梳離肺組織 ，直到肺組織與支氣管樹全部分離。使肺組織通過00目的鋼絲網 ，分離單細胞。4、以下過程均在4℃中進行。
Isolation and Purification of Alveolar Macrophages from Rabbits Macrophage Isolation and Purification 1. Rabbits weighing 2 to 3 kilograms were killed by intravenous injection of 10 ml of air into the ear vein or injected with 2 ml (containing 130 mg) of pentobarbital anesthetized animals. . Supine fixation, disinfection, cut the chest wall. The following procedure pays attention to aseptic operation. Carefully cut the trachea from beneath the cricoid cartilage, separate the heart and blood vessels, remove the lungs, and cut fat and connective tissue. The lung surface was carefully wiped with sterile gauze and weighed. 2. Detach the alveolar macrophage: clip the trachea with the clip and let the lungs float. Into the trachea, 40 ml of sterile cold saline was injected, and physiological saline was allowed to enter all alveoli. Pull the trachea with forceps, cut the trachea from underneath the clamp, and pour the lung fluid into the centrifuge tube. In the same manner, the alveoli were washed with 40 ml of sterile saline, and the combined leachate was set at 4°C. 3. To isolate macrophages from lung tissue: After removing the alveolar macrophages, cut the trachea and place the lung tissue in a plate with cold RPMI-1640 medium. The dish was placed on ice, and a 23-gauge needle was connected to a 20-ml syringe that was filled with cold RPMI-1640 medium, and the needle was used to perfuse the lung tissue until the lung tissues were compley separated from the bronchial tree. The lung tissue was passed through a 00 mesh wire mesh to separate single cells. 4. The following procedures were all performed at 4°C..