Sphere-Formation Assay: Three-Dimensional in vitro Culturing of Prostate Cancer Stem/Progenitor Sphere-Forming Cells.
Sphere-Formation Assay: Three-Dimensional in vitro Culturing of Prostate Cancer Stem/Progenitor Sphere-Forming Cells.
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DOI：
10.3389/fonc.2018.00347
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发表时间：
2018
影响因子：
4.7
通讯作者：
Abou-Kheir W
中科院分区：
文献类型：
--
作者：
Bahmad HF;Cheaito K;Chalhoub RM;Hadadeh O;Monzer A;Ballout F;El-Hajj A;Mukherji D;Liu YN;Daoud G;Abou-Kheir W
癌症干细胞（CSC）是在大多数肿瘤中鉴定的细胞亚群，负责不同恶性肿瘤的起始、复发、转移潜力和抗性。在前列腺癌（PCa）中，CSC被鉴定并被认为是导致致死亚型（通常称为去势抵抗性前列腺癌（CRPC））产生的原因。研究PCa中CSC特性的体外模型是非常必要的。成球试验是一种常用的体外鉴定CSCs并研究其特性的方法。在这里，我们报告了如何从前列腺癌细胞系和小鼠前列腺组织中产生和繁殖球体的详细方法。该模型基于干细胞在非粘附无血清凝胶基质中生长的能力。我们还描述了如何在组织学和免疫荧光染色测定中使用这些球体来评估CSC的分化潜力。我们的研究结果表明，球形成试验（SFA）作为一个可靠的体外试验，以评估在不同的PCa模型中的CSC的存在和自我更新能力。该平台提供了一个有用的工具来评估传统或新型药物对不同肿瘤的启动和自我更新特性的影响。可以通过评估五代以上的球体形成效率（SFE）或其他下游测定（例如对所产生的球体的免疫组织化学分析）来直接评估效果。
Cancer Stem Cells (CSCs) are a sub-population of cells, identified in most tumors, responsible for the initiation, recurrence, metastatic potential, and resistance of different malignancies. In prostate cancer (PCa), CSCs were identified and thought to be responsible for the generation of the lethal subtype, commonly known as Castration-Resistant Prostate Cancer (CRPC). In vitro models to investigate the properties of CSCs in PCa are highly required. Sphere-formation assay is an in vitro method commonly used to identify CSCs and study their properties. Here, we report the detailed methodology on how to generate and propagate spheres from PCa cell lines and from murine prostate tissue. This model is based on the ability of stem cells to grow in non-adherent serum-free gel matrix. We also describe how to use these spheres in histological and immuno-fluorescent staining assays to assess the differentiation potential of the CSCs. Our results show the sphere-formation Assay (SFA) as a reliable in vitro assay to assess the presence and self-renewal ability of CSCs in different PCa models. This platform presents a useful tool to evaluate the effect of conventional or novel agents on the initiation and self-renewing properties of different tumors. The effects can be directly evaluated through assessment of the sphere-forming efficiency (SFE) over five generations or other downstream assays such as immuno-histochemical analysis of the generated spheres.
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影响因子：
10.5
作者：
Dontu, G;Abdallah, WM;Wicha, MS
通讯作者：
Wicha, MS
影响因子：
--
作者：
Daoud G;Monzer A;Bahmad H;Chamaa F;Hamdar L;Mouhieddine TH;Shayya S;Eid A;Kobeissy F;Liu YN;Abou-Kheir W
通讯作者：
Abou-Kheir W
影响因子：
2.8
作者：
Garraway, Isla P.;Sun, Wenyi;Tran, Chau P.;Perner, Sven;Zhang, Bao;Goldstein, Andrew S.;Hahm, Scott A.;Haider, Maahum;Head, Christian S.;Reiter, Robert E.;Rubin, Mark A.;Witte, Owen N.
通讯作者：
Witte, Owen N.
影响因子：
1.2
作者：
Dong, Qiaoxiang;Wang, Danhan;Bandyopadhyay, Abhik;Gao, Hui;Gorena, Karla M.;Hildreth, Kim;Rebel, Vivienne I.;Walter, Christi A.;Huang, Changjiang;Sun, Lu-Zhe
通讯作者：
Sun, Lu-Zhe
影响因子：
6.6
作者：
Goldstein AS;Stoyanova T;Witte ON
通讯作者：
Witte ON