Single-cell imaging of human cancer xenografts using adult immunodeficient zebrafish.
Single-cell imaging of human cancer xenografts using adult immunodeficient zebrafish.
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DOI：
10.1038/s41596-020-0372-y
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发表时间：
2020-09
期刊：
影响因子：
14.8
通讯作者：
Langenau DM
中科院分区：
文献类型：
--
作者：
Yan C;Do D;Yang Q;Brunson DC;Rawls JF;Langenau DM
斑马鱼是一种理想的细胞移植模型。它们繁殖力强，光学透明，是临床前药物发现研究的绝佳平台。传统上，异种移植是使用尚未发育出适应性免疫的幼斑马鱼进行的。幼虫移植是一个强大的短期移植平台，适合高通量药物筛选研究，但动物最终会排斥肿瘤，不能在37 °C下饲养。为了解决这些局限性，我们最近开发了成年casper品系prkdc−/−，il 2 rg α−/−免疫受损的斑马鱼，这些斑马鱼能够稳健地植入人类癌细胞超过28天。由于成年斑马鱼可以通过口服灌胃或腹腔注射给药，因此我们的模型适用于实现准确的临床前药物给药。我们的平台还允许在几天内以单细胞分辨率轻松可视化体内药物作用。在这里，我们描述了将异种移植细胞移植到prkdc−/−，il 2 rg α−/−模型中的程序，包括在37 °C下优化免疫抑制斑马鱼生长和饲养的精细饲养方案;优化的腹膜内和眼周肌细胞移植;以及荧光和共聚焦成像方法，以体内单细胞分辨率可视化给予临床相关药物剂量的效果。在鉴定成年纯合子动物后，该程序需要35天才能完成。使成鱼适应37°C需要7天，植入研究需要28天。我们的方案为使用免疫受损的斑马鱼进行异种移植细胞移植提供了全面的指导，并将该模型作为一种新的临床前药物发现平台。该方案描述了如何将人类癌细胞移植到免疫受损的成年斑马鱼中。首先使鱼适应37°C，然后进行异种移植细胞的腹膜内或眼周肌肉移植和荧光成像。
Zebrafish are an ideal cell transplantation model. They are highly fecund, optically clear and an excellent platform for preclinical drug discovery studies. Traditionally, xenotransplantation has been carried out using larval zebrafish that have not yet developed adaptive immunity. Larval engraftment is a powerful short-term transplant platform amenable to high-throughput drug screening studies, yet animals eventually reject tumors and cannot be raised at 37 °C. To address these limitations, we have recently developed adult casper-strain prkdc−/−, il2rgα−/− immune compromised zebrafish that robustly engraft human cancer cells for in excess of 28 days. Because the adult zebrafish can be administered drugs by oral gavage or intraperitoneal injection, our model is suitable for achieving accurate, preclinical drug dosing. Our platform also allows facile visualization of drug effects in vivo at single cell resolution over days. Here, we describe the procedures for xenograft cell transplantation into the prkdc−/−, il2rgα−/− model, including refined husbandry protocols for optimal growth and rearing of immune suppressed zebrafish at 37 °C; optimized intraperitoneal and periocular muscle cell transplantation; and epifluorescence and confocal imaging approaches to visualize the effects of administering clinically relevant drug dosing at single cell resolution in vivo. Following identification of adult homozygous animals, this procedure takes 35 days to complete. 7 days are required to acclimate adult fish to 37°C and 28 days are required for engraftment studies. Our protocol provides a comprehensive guide for using immune compromised zebrafish for xenograft cell transplantation and credentials the model as a new preclinical drug discovery platform. This protocol describes how to engraft human cancer cells in immune compromised adult zebrafish. The fish are first adapted to 37°C, followed by intraperitoneal or periocular muscle transplantation of xenograft cells and fluorescence imaging.
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染色体不稳定性通过胞质DNA反应驱动转移。
影响因子：
64.8
作者：
Bakhoum SF;Ngo B;Laughney AM;Cavallo JA;Murphy CJ;Ly P;Shah P;Sriram RK;Watkins TBK;Taunk NK;Duran M;Pauli C;Shaw C;Chadalavada K;Rajasekhar VK;Genovese G;Venkatesan S;Birkbak NJ;McGranahan N;Lundquist M;LaPlant Q;Healey JH;Elemento O;Chung CH;Lee NY;Imielenski M;Nanjangud G;Pe'er D;Cleveland DW;Powell SN;Lammerding J;Swanton C;Cantley LC
通讯作者：
Cantley LC
影响因子：
64.8
作者：
Stewart E;Federico SM;Chen X;Shelat AA;Bradley C;Gordon B;Karlstrom A;Twarog NR;Clay MR;Bahrami A;Freeman BB 3rd;Xu B;Zhou X;Wu J;Honnell V;Ocarz M;Blankenship K;Dapper J;Mardis ER;Wilson RK;Downing J;Zhang J;Easton J;Pappo A;Dyer MA
通讯作者：
Dyer MA
影响因子：
2.9
作者：
Pereiro, P.;Varela, M.;Novoa, B.
通讯作者：
Novoa, B.
影响因子：
28.2
作者：
Drapkin BJ;George J;Christensen CL;Mino-Kenudson M;Dries R;Sundaresan T;Phat S;Myers DT;Zhong J;Igo P;Hazar-Rethinam MH;Licausi JA;Gomez-Caraballo M;Kem M;Jani KN;Azimi R;Abedpour N;Menon R;Lakis S;Heist RS;Büttner R;Haas S;Sequist LV;Shaw AT;Wong KK;Hata AN;Toner M;Maheswaran S;Haber DA;Peifer M;Dyson N;Thomas RK;Farago AF
通讯作者：
Farago AF
影响因子：
3.8
作者：
Cabezas-Sainz, Pablo;Guerra-Varela, Jorge;Sanchez, Laura
通讯作者：
Sanchez, Laura