Cancer Research
A subscription to JoVE is required to view this content.
You will only be able to see the first 2 minutes.
The JoVE video player is compatible with HTML5 and Adobe Flash. Older browsers that do not support HTML5 and the H.264 video codec will still use a Flash-based video player. We recommend downloading the newest version of Flash here, but we support all versions 10 and above.
If that doesn't help, please let us know.
为癌症药理学研究创建匹配的体内/体外患者衍生模型对 PDX 和 PDX 衍生器官
Chapters
Summary May 5th, 2021
Please note that all translations are automatically generated.
Click here for the English version.
描述使用患者衍生的异种格拉夫特 （PDX） 进行体外筛查的方法，从而形成匹配的体内/体外模型对。PDX 肿瘤被机械或酶地收获/加工成小块，然后是 Clevers 的生长肿瘤器官的方法，这些器官是通过、冷冻保存和针对原始 PDX 进行特征的。
Transcript
此协议可用于建立配对患者衍生的异种移植物（PDX）和相应的患者衍生器官线的匹配库。这种方法的优点是，它可以用来创建使用PDX进行体外筛查的器官，从而产生匹配的体外/体外模型对。首先收获肿瘤并处理文本手稿中描述的组织。
组织消化后，用五毫升无菌塑料移液器上下吹笛。然后将 20 毫升 AD+添加到同质化中，然后用 100 微米细胞过滤器进行过滤。用 AD+清洗两次，然后将其转移到塑料管中，然后以 450 倍 G 的离心处理，为期五分钟。
将 BME 中的细胞重新保存并保存在冰上。为了准备器官培养，将细胞悬浮物的200微升转移到六井板的每口井中，并在37摄氏度下孵育板30分钟。在每口井中加入两毫升的有机介质，并图像代表在显微镜下滴落。
将器官培养物保持在37摄氏度和5%的二氧化碳中，每三到四天发生一次中等变化，每七天以一比二的比例通过。分离器官后，通过 70 微米滤镜运行到 50 毫升塑料管中。在显微镜下数一数器官，并在冰上将其重新注入BME，最终浓度为5%，在每384井板中加入50微升的器官悬浮剂，并配有液体分配器，每口油井的播种密度为200 CR2110 PDXO。
使用数字分配器在串行稀释中将 SN38 添加到每个油井中。使用数字分配器软件工具创建板图。治疗应包括负控制车辆与100%的生存能力和积极控制五微摩尔葡萄球菌素与0%的可行性。
完成后，将经过药物处理的384井板放回37摄氏度的孵化器中。在光显微镜下，PDXO CR2110 显示了典型的囊性形态，表明 PDX 衍生器官与患者衍生器官在相同培养条件下的相似性。H&E染色组织的病理学检查表明，PDXO CR2110的组织结构和细胞类型与原始的PDX CR2110相似。
PDX 和 PDXO 的基因组特征比较表明转录体表达的相关性为 94.92%，DNA 突变的一致性为 97.67%，这表明这两个模型之间的整体基因组相似性。在384井板的PDXO CR2110上进行了药物敏感性检测。PDXO CR2110对伊里诺特坎敏感，对西斯铂具有抗药性，与PDX治疗结果一致。
匹配的体外模型和体内模型可以互相称赞体外筛查和体内验证，提高药物发现的成功率，并可能降低临床开发中的自然减员率。
Related Videos
You might already have access to this content!
Please enter your Institution or Company email below to check.
has access to
Please create a free JoVE account to get access
Login to access JoVE
Please login to your JoVE account to get access
We use/store this info to ensure you have proper access and that your account is secure. We may use this info to send you notifications about your account, your institutional access, and/or other related products. To learn more about our GDPR policies click here.
If you want more info regarding data storage, please contact gdpr@jove.com.
Please enter your email address so we may send you a link to reset your password.
We use/store this info to ensure you have proper access and that your account is secure. We may use this info to send you notifications about your account, your institutional access, and/or other related products. To learn more about our GDPR policies click here.
If you want more info regarding data storage, please contact gdpr@jove.com.
Your JoVE Unlimited Free Trial
Fill the form to request your free trial.
We use/store this info to ensure you have proper access and that your account is secure. We may use this info to send you notifications about your account, your institutional access, and/or other related products. To learn more about our GDPR policies click here.
If you want more info regarding data storage, please contact gdpr@jove.com.
Thank You!
A JoVE representative will be in touch with you shortly.
Thank You!
You have already requested a trial and a JoVE representative will be in touch with you shortly. If you need immediate assistance, please email us at subscriptions@jove.com.
Thank You!
Please enjoy a free 2-hour trial. In order to begin, please login.
Thank You!
You have unlocked a 2-hour free trial now. All JoVE videos and articles can be accessed for free.
To get started, a verification email has been sent to email@institution.com. Please follow the link in the email to activate your free trial account. If you do not see the message in your inbox, please check your "Spam" folder.
