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1 实验步骤、方法、注意事项、出现问题的原因 C_(57)BL/6于右侧腋下皮下接种Lewis肺癌瘤株,待肿瘤长至1.5cm×1.5cm,在超净工作台无菌剥离,置入HBSS液中,剪取边缘新鲜瘤组织成1nm× 1mm,置于RPMI-1640配制的消化液中稍加改良, DNase Ⅰ 0.1mg/ml,collagenase Ⅳ 1mg/ml, hyaluronidase:0.5mg/10ml, 于37℃孵箱消化4 h,经120目尼龙网过滤,离心1500r/min,5min弃上清;用HBSS液洗一次,弃上清,加入约2mlHBSS,吹打制成细胞悬液,准备细胞分离。
1 Experimental procedures, methods, precautions, reasons for the problem C_(57)BL/6 was inoculated subcutaneously in the right armpit with Lewis lung cancer tumor until the tumor grew to 1.5 cm × 1.5 cm, and was aseptically stripped at the clean bench. Place the HBSS solution, cut the edge of the fresh tumor tissue into 1nm × 1mm, placed slightly modified in the RPMI-1640 preparation of the digestive juice, DNase I 0.1mg/ml, collagenase IV 1mg/ml, hyaluronidase: 0.5mg/ 10 ml, digested at 37°C for 4 h, filtered through 120 mesh nylon mesh, centrifuged at 1500 r/min, and discarded for 5 min; supernatant was washed with HBSS once, supernatant was discarded, approximately 2 ml of HBSS was added, and cell suspension was prepared. Cell separation.