兔 vx2 模型在肝癌中的开发与应用
Summary
本文的目的是为 vx2 肝癌兔模型的开发和应用提供一个入门。
Abstract
兔 vx2 肿瘤是介入放射学领域肝细胞癌 (hcc) 转化研究的常用动物模型。该模型采用了一种再生鳞状细胞癌, 该细胞癌在供体家兔的骨骼肌中容易和可靠地传播, 以便最终收获并同种异体移植植入天真受者的肝脏。这种肿瘤移植在受者兔的肝脏中迅速生长成血管造影可识别的肿瘤, 其特征是由一个可行的血管高血管囊包围的坏死核。兔子解剖的物理大小足以促进血管器械, 允许各种介入技术的应用和测试。尽管有这些好处, 但缺乏技术资源, 无法作为研究人员使用该模型的具体参考。在此, 我们提出了一个全面的视觉概述的技术方面的发展, 生长, 繁殖, 和血管造影利用的兔子 vx2 肿瘤模型, 供新手和有经验的研究人员使用。
Introduction
兔 vx2 肿瘤模型自 1935年 1,2发展以来, 在实验肿瘤学中发挥了重要作用。该肿瘤是一种病毒引起的再生鳞状细胞癌, 其特征是血管增生、生长快、在骨骼肌中容易繁殖 3,4。而兔 vx2 肿瘤模型已被用来研究多种癌症5,6,7,8;本文的重点是肝癌9。
所述方法的目的是提出一个模型的原发性肝癌, 或肝细胞癌 (hcc), 可用于介入放射科医生的翻译研究。它可用于药代动力学研究、治疗调查和消融方法检测10,11,12,13,14,15.
与老鼠、老鼠、土木等老鼠模型或灵长类动物 1 6号等较大模型等其他模型相比, 本文详细介绍的方法具有多重优势.主要好处之一是肿瘤生长迅速可靠, 这使得研究人员能够在第一次后肢繁殖后一个月内建立活跃的肿瘤线.此外, 这种肿瘤有直接的超声表现可见性和高血管的外围, 这允许跨动脉局部治疗和消融治疗。最后, 也是最重要的, 兔子血管的大小允许可行和技术上容易利用血管器械 18.
Protocol
以下协议遵循伊利诺伊大学芝加哥分校规定的所有要求和准则。它在执行前经过了当地动物护理和使用机构委员会的审查和批准。 1. vx2 hind 肢体肿瘤的发展 从国家癌症研究所癌症治疗诊断和治疗肿瘤细胞系存储库中获得 vx2 肿瘤细胞系。注意: 此时, 订单目录可在以下链接中找到: https://dtp.cancer.gov/repositories/。 准备注射用细胞悬浮液, 将冷冻的 vx2 样品和甲基…
Representative Results
当看图 1, 很明显, 兔子的四头肌扩大。此外, 多个小的离散结节, 通常与肿瘤生长通过筋膜相关, 是可见的。触诊时, 注射肢体应出现比非注射肢体。如果研究人员需要更明确的肿瘤存在保证, 超声成像可以用来识别嵌入肌肉中的肿瘤。如果没有检测到肿瘤, 后肢应重新注射肿瘤细胞悬浮液。 为了确认成功?…
Discussion
vx2 肿瘤方法的第一个关键步骤是成功地在供体兔的后肢传播肿瘤。有关此步骤的详细信息, 请参阅 “代表性结果” 部分的第一段。
下一个关键步骤是确保正确识别可行的肿瘤胶囊。这不仅对肿瘤悬浮液的制备是必要的, 对肝植入的肿瘤碎片的选择和产生也很重要。图 4对活肿瘤和周围肌肉组织之间的界限进行了注释。如果在悬浮液制备过程中刮伤不正确的…
Divulgations
The authors have nothing to disclose.
Acknowledgements
我们要感谢伊利诺伊大学芝加哥生物资源实验室的兽医工作人员。
Materials
| MethoCult (Methycellulose) | Stemcell Technologies | M3134 | |
| VX2 Cell Line | NCI | VX-2 | |
| 5 mL Syringe | BD | 309646 | |
| 16-Gauge Needle | BD | 305197 | |
| 22-Gauge Needle | BD | 305155 | |
| Hair Clippers | Wahl | 41870-0438 | |
| Foam Insulated Box | Mr. Box Online | 10 x 10 x 4 | |
| Acepromazine | Henry Schein | 003845 | |
| Buprenorphine | Par | 42023-179-05 | |
| Meloxicam | Henry Schein | 049755 | |
| Alcohol Pads | Covidien | 5033 | |
| Ketamine | Henry Schein | 056344 | |
| Xylazine | Akorn | 59399-110-20 | |
| Pentobarbital (Fatal-Plus) | Vortech | 9373 | |
| Sterile Petri Dish | Thermo Fisher | 172931 | |
| DMEM | Gibco | 11965092 | |
| Saline | Baxter | 2F7124 | |
| 15-Blade | Steris | 02-050-015 | |
| Scalpel Handle x 2 | Steris | 22-2381 | |
| Curved Hemostat | WPI | 501288 | |
| Atraumatic Forceps | Sklar | 52-5077 | |
| Gauze | Medline | NON21430LF | |
| 11-Blade | Steris | 02-050-011 | |
| Surgicel | Ethicon | 1951 | |
| 3-0 PDS / Taper | Ethicon | Z305H | |
| 4 – 0 Vicryl / Cutting | Ethicon | J392H | |
| 40 micron strainer | BD | 352340 | |
| 50 mL conical tube | Thermo Fisher | 339652 | |
| plastic pipette | Thomas Scientific | HS206371B | |
| Centrifuge | Sorvall | 75004240 | |
| 1.40mL Tubes (Internal Thread) | Micronic | MP32131-Z20 | |
| 3-F VSI Micro-HV Introducer Kit | Vascular Solutions | Custom Order (P15180391) | |
| .018 45-degree angle glidewire | Terumo | RG*GA1818SA | |
| Direxion bern-shape microcatheter | Boston Scientific | M001195230 | |
| Omnipaque | GE | Y510 |
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Citer Cet Article
Khabbaz, R. C., Huang, Y., Smith, A. A., Garcia, K. D., Lokken, R. P., Gaba, R. C. Development and Angiographic Use of the Rabbit VX2 Model for Liver Cancer. J. Vis. Exp. (143), e58600, doi:10.3791/58600 (2019).