细胞系特征 |
细胞株名称:A549/DDP人肺癌顺铂耐药株 种属:人 组织来源:肺癌 生长特性:贴壁生长 形态特征:上皮细胞 微生物及支原体检测:阴性 安全性:所有肿瘤和病毒转染的细胞均视为有潜在的生物危害性,必须在二级生物安全台内操作,并请注意防护,所有废液及接触过此细胞的器皿需高压灭菌后方能丢弃。 |
培养条件: |
完全培养基:90%RPMI-1640+10%胎牛血清+2000ng/ml DDP 血清我们推荐用: GIBCOFBS-10099-141或HYCLONEFBS-SH30084.03。 培养条件:37.0C carbon dioxide(CO2),5% |
传代方法: |
收到细胞后,在倒置镜下(最好是在4X物镜)观察整个细胞生长情况。 (一)如果细胞未长满,用75%酒精喷洒整个瓶消毒后放到超菌台内,严格无菌操作,打开细胞培养瓶,吸出培养液,换 10ml新鲜培养液后继续培养。 (二)如果细胞已长满,即可进行传代培养。具体步骤如下: 1. 弃去培养液,用PBS(不含钙,镁离子)洗1-2次。 2. 加0.7-1ml消化液(0.25%Trypsin-0.53mM EDTA)于培养瓶中,用力拍打瓶壁,期间每隔 5-10s放到显微镜下观察,直至50-70%的细胞脱落后,加入2ml 以上完全培养基中止消化。 3. 按6-8ml/瓶补加完全培养基,轻轻打匀后吸出一半,分到新的培养瓶中。如果没有特别说明,收到细胞后的第一次传代一般是一传二。 注:1、观察细胞最好在低倍镜(4或5X物镜)下进行,否则不能准确判断细胞的传代密度。看细胞的形态请在10X或20X物镜下。 2、瓶中运输培养基不能重复再用,请换用加双抗的新培养基,细胞冻存后,培养基中可不加任何抗生素。 3、有些细胞贴壁不牢,如发现贴壁细胞有脱落,可离心吹打后接种到新瓶内。 4、收到细胞后,若发现培养瓶破损、有液溢出及细胞有污染,请及时与我们联系。.. |
冻存方法: |
冻存液:90%胎牛血清,10%DMSO 储存:液氮储存 |
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apoptosis, marked by greater caspase-9 expression [6, 78, 80]. In addition, in a Beclin-1-
deficient T cell model, decreased numbers of CD4+ and CD8+ T cells correlated with
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