Conditional reprogramming and long-term expansion of normal and tumor cells from human biospecimens.
Conditional reprogramming and long-term expansion of normal and tumor cells from human biospecimens.
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DOI：
10.1038/nprot.2016.174
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发表时间：
2017-03
期刊：
影响因子：
14.8
通讯作者：
Schlegel R
中科院分区：
文献类型：
--
作者：
Liu X;Krawczyk E;Suprynowicz FA;Palechor-Ceron N;Yuan H;Dakic A;Simic V;Zheng YL;Sripadhan P;Chen C;Lu J;Hou TW;Choudhury S;Kallakury B;Tang DG;Darling T;Thangapazham R;Timofeeva O;Dritschilo A;Randell SH;Albanese C;Agarwal S;Schlegel R
从历史上看，直接从人类肿瘤或健康组织中获得的细胞很难在体外繁殖。然而，体外临床前模型是基础癌症生物学研究和促进转化研究（包括药物发现和药物靶点鉴定）的重要工具。该方案描述了条件重编程（CR），包括在Rho激酶抑制剂（Y-27632）存在下，将辐照小鼠成纤维细胞饲养细胞与正常和肿瘤人上皮细胞共培养。CR细胞可用于各种应用，包括再生医学，药物敏感性测试，基因表达谱和异种移植研究。该方法需要病理学家区分健康组织和肿瘤组织，以及基本的组织培养技能。该方案可用于来自新鲜和冷冻保存的组织样本的细胞。由于在7天内可以产生大约100万个细胞，该技术可以直接应用于诊断和预测医学。此外，上皮细胞可以在体外无限繁殖，但在模拟其自然环境的条件下保留完全分化的能力。
Historically, it has been difficult to propagate cells in vitro that are derived directly from human tumors or healthy tissue. However, in vitro preclinical models are essential tools for both the study of basic cancer biology and the promotion of translational research, including drug discovery and drug target identification. This protocol describes conditional reprogramming (CR), which involves coculture of irradiated mouse fibroblast feeder cells with normal and tumor human epithelial cells in the presence of a Rho kinase inhibitor (Y-27632). CR cells can be used for various applications, including regenerative medicine, drug sensitivity testing, gene expression profiling and xenograft studies. The method requires a pathologist to differentiate healthy tissue from tumor tissue, and basic tissue culture skills. The protocol can be used with cells derived from both fresh and cryopreserved tissue samples. As approximately 1 million cells can be generated in 7 d, the technique is directly applicable to diagnostic and predictive medicine. Moreover, the epithelial cells can be propagated indefinitely in vitro, yet retain the capacity to become fully differentiated when placed into conditions that mimic their natural environment.
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影响因子：
12.3
作者：
Nadauld LD;Garcia S;Natsoulis G;Bell JM;Miotke L;Hopmans ES;Xu H;Pai RK;Palm C;Regan JF;Chen H;Flaherty P;Ootani A;Zhang NR;Ford JM;Kuo CJ;Ji HP
通讯作者：
Ji HP
影响因子：
3.8
作者：
Brown, Daniel D.;Dabbs, David J.;McAuliffe, Priscilla F.
通讯作者：
McAuliffe, Priscilla F.
影响因子：
8
作者：
Charette, S. T.;McCance, D. J.
通讯作者：
McCance, D. J.
影响因子：
6
作者：
Palechor-Ceron, Nancy;Suprynowicz, Frank A.;Liu, Xuefeng
通讯作者：
Liu, Xuefeng
影响因子：
82.9
作者：
Hata AN;Niederst MJ;Archibald HL;Gomez-Caraballo M;Siddiqui FM;Mulvey HE;Maruvka YE;Ji F;Bhang HE;Krishnamurthy Radhakrishna V;Siravegna G;Hu H;Raoof S;Lockerman E;Kalsy A;Lee D;Keating CL;Ruddy DA;Damon LJ;Crystal AS;Costa C;Piotrowska Z;Bardelli A;Iafrate AJ;Sadreyev RI;Stegmeier F;Getz G;Sequist LV;Faber AC;Engelman JA
通讯作者：
Engelman JA