細胞培養 RPMI Medium 1640

平時研究用的細胞，有需要冷凍保存。冷凍的過程稱為凍存，把凍上的細胞化開的過程叫細胞復蘇。細胞復蘇是一簡單的試驗操作，但操作中有許多需要注意的事項，在實驗操作中注意細小問題，才能使復蘇后的細胞狀態保持良好。

細胞復蘇實驗準備：

主要器材：一次性滴管、打火機、酒精燈、大鑷子、中鑷子、記號筆、廢液缸、封口膜、剪子。

主要試劑： PBS、培養液、消化酶（胰酶）、75%酒精、雙抗。

PBS配制：NaCL :4g   KCL:0.1g   Na2HPO4 :1.745g   KH2PO4 :0.1g   

三蒸水： 500mL，溶解后高壓滅菌。

培養基配制：5mL雙抗、50mL血清，加入培養基中。

細胞復蘇操作步驟：

1.佩戴眼鏡和手套，從液氮罐中取出安瓿或冷凍管。

2.迅速放入38℃水浴中，并不時搖動，在1分鐘內使其*融化，然后在無菌下取出細胞。

3.在1000r/min速度下離心5～10分鐘，棄去上層液，加入適量培養液后接種于培養瓶中，接種濃度1×109/L，置37℃溫箱靜置培養，次日更換一次培養液，繼續培養，觀察生長情況。若細胞密度較高，及時傳代。或無需離心直接將細胞加入瓶中，并加入培養基貼壁培養12～24小時后，充去上清，換入新鮮培養基繼續培養。

相關文獻：

《c-Myc induced upregulation of long non-coding RNA SNHG16 enhances progression and carcinogenesis in oral squamous cell carcinoma》 作者:Shangfeng Li, Shengkai Zhang， Jie Chen 期刊:Cancer gene therapy 影響因子:4.681 PMID:30607006
《Suberoylanilide hydroxamic acid inhibits proliferation and promotes apoptosis of mouse myeloma Sp2/0 cells through activating p53/caspase-3 signaling》 作者:Yanliang Bai, Lei Huo, Chenyu Zhang, Mingyue Shi, Na Fang, Shaoping Ji, Kai Sun 期刊:INTERNATIONAL JOURNAL OF CLINICAL AND EXPERIMENTAL MEDICINE 影響因子:1.875 PMID:
《Gecko Active Components Regulates Endoplasmic Reticulum Stress to Induce the Apoptosis of KYSE150 Cells through PERK Pathway》 作者:Yimeng Duan, Lengxin Duan, Zeyue Huang, Bingbing Wang, Jiangang Wang 期刊:Journal of Biosciences and Life Sciences 影響因子:0.552 PMID:
《RNA-binding protein RBM6 as a tumor suppressor gene represses the growth and progression in laryngocarcinoma》 作者:Qiwei Wang,Fang Wang,Waisheng Zhong,Hang Ling,Jixuan Wang,Jie Cui,Tao Xie,Senli Wen,Jie Chen 期刊:Gene 影響因子:2.638 PMID:30772516
《Sphingomyelin synthase 2 promotes H2O2-induced endothelial dysfunction by activating the Wnt/β-catenin signaling pathway》 作者:Panpan Zhang,Lingyue Hua,Huan Hou, Xingyue Du,Zhiqiang He, Menghan Liu, Xiaojuan Hu, Nianlong Yan 期刊:International Journal of Molecular Medicine 影響因子:2.928 PMID:30272329
《Downregulation of transcription factor EB inhibits the growth and metastasis of colorectal carcinomas》 作者:Jin-Zhe Chang,Shu-Dong Chen,Hui Zheng,Hua-Ping Zhang 期刊:European Journal of Inflammation 影響因子:0.733 PMID:
《Circular RNA hsa_circ_0002024 suppresses cell proliferation, migration, and invasion in bladder cancer by sponging miR-197-3p》 作者:Yongjun Jiang, Tingran Wei, Wei Li, Ruihua Zhang, and Maogang Chen 期刊:American journal of Translational Research 影響因子:3.266 PMID:30972190
《Effect of microRNA‐128 on cisplatin resistance of glioma SHG‐44 cells by targeting JAG1》 作者:Dong‐Ye Yi， Qing Su， Fang‐Cheng Zhang， Peng Fu， Qing Zhang， Yong‐Cun Cen， Hong‐Yang Zhao，Wei Xiang 期刊:Journal of Cellular Biochemistry 影響因子:3.448 PMID:29091297
《miR‐377‐5p inhibits lung cancer cell proliferation, invasion, and cell cycle progression by targeting AKT1 signaling》 作者:Han Wu，Haiyan Liu,Wenjie Liu,Yongli Shi,Dawei Bao 期刊:Journal of Cellular Biochemistry 影響因子:3.448 PMID:30485528
《Activity of coumarin against Candida albicans biofilms》 作者:K.Xu  J.L.Wang  M.P.Chu  C.Jia 期刊:Journal de Mycologie Medicale 影響因子:1.479 PMID:30606640
《Gallic acid has anticancer activity and enhances the anticancer effects of cisplatin in non?small cell lung cancer A549 cells via the JAK/STAT3 signaling pathway》 作者:Tingxiu Zhang, Lijie Ma, Pengfei Wu, Wei Li, Ting Li, Rui Gu, Xiaoping Dan, Zhiwei Li, Xianming Fan, Zhenliang Xiao 期刊:Oncology Reports 影響因子:3.041 PMID:30747218
《Downregulation of lncRNA NEAT1_2 radiosensitizes hepatocellular carcinoma cells through regulation of miR‐101‐3p/WEE1 axis》 作者:Xin Chen, Nuobei Zhang 期刊:Cell Biology International 影響因子:2.127 PMID:30488993
《Knockdown of USP28 enhances the radiosensitivity of esophageal cancer cells via the c‐Myc/hypoxia‐inducible factor‐1 alpha pathway》 作者:Zhang Weili， Liu Zhikun，Wang Jianmin， Tian Qingbao 期刊:Journal of Cellular Biochemistry 影響因子:3.448 PMID:30206969
《TINAGL1 promotes hepatocellular carcinogenesis through the activation of TGF-β signaling-medicated VEGF expression》 作者:Lu Sun, Zihui Dong, Hongli Gu, Zhixian Guo, and Zujiang Yu 期刊:Cancer Management and Research 影響因子:2.243 PMID:30697069
《LPS?induced proinflammatory cytokine expression in human airway epithelial cells and macrophages via NF?κB, STAT3 or AP?1 activation》 作者:Xuefang Liu， Sugai Yin， Yulong Chen， Yaosong Wu， Wanchun Zheng， Haoran Dong， Yan Bai， Yanqin Qin， Jiansheng Li， Suxiang Feng， Peng Zhao 期刊:Molecular Medicine Reports 影響因子:1.851 PMID:29393460
《A Host Factor GPNMB Restricts Porcine Circovirus Type 2 (PCV2) Replication and Interacts With PCV2 ORF5 Protein》 作者:Kangkang Guo, Lei Xu, Mengmeng Wu, Yufeng Hou, Yanfen Jiang, Jiangman Lv, Panpan Xu, Zhixin Fan, Ruiqi Zhang, Fushan Xing and Yanming Zhang 期刊:Frontiers in Microbiology 影響因子:4.019 PMID:30671053

細胞培養 RPMI Medium 1640