RNAseq in lung cancer pneumospheres treated with PD-L1s. RNAseq in lung cancer pneumospheres treated with PD-L1s

We evaluated the differential expression of protein coding genes in NSCLC pneumospheres upon treatment with PD-L1s Overall design: Pneumospheres were generated seeding NSCLC cells in Ultra-Low Attachment flasks, in serum free DMEM-F12 medium containing fibroblast growth factor, epithelial growth factor and B-27™ Supplement. NSCLC pneumospheres were stimulated by synthetic PD-L1 soluble form (PD-L1s) (25 μg/ml, R&D system) in 0% FBS culture medium for 15 min.本研究评估了经PD-L1s处理后，非小细胞肺癌（Non-Small Cell Lung Cancer, NSCLC）肺球体中蛋白编码基因的差异表达情况。整体实验设计：肺球体的构建方法为：将非小细胞肺癌细胞接种于超低吸附培养瓶内，使用添加了成纤维细胞生长因子、表皮生长因子及B-27™添加剂的无血清DMEM-F12培养基进行培养。将上述非小细胞肺癌肺球体置于0%胎牛血清培养基中，以浓度为25 μg/mL的合成可溶性PD-L1（PD-L1s，R&D系统公司产品）刺激处理15分钟。