Production of Uniform 3D Microtumors in Hydrogel Microwell Arrays for Measurement of Viability, Morphology, and Signaling Pathway Activation.
Production of Uniform 3D Microtumors in Hydrogel Microwell Arrays for Measurement of Viability, Morphology, and Signaling Pathway Activation.
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在水凝胶微孔阵列中生产均匀的 3D 微肿瘤，用于测量活力、形态和信号通路激活。
DOI：
10.1089/adt.2015.662
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发表时间：
2015
影响因子：
1.8
通讯作者：
Sant,Shilpa
中科院分区：
文献类型：
--
作者：
Singh,Manjulata;Close,DavidA;Mukundan,Shilpaa;Johnston,PaulA;Sant,Shilpa
尽管在癌症研究和药物发现/开发方面投入了大量资金，但新癌症药物的批准率≤5%，大多数转移性癌症病例仍然无法治愈。95%的癌症新药在临床开发中失败，因为缺乏治疗效果和/或不可接受的毒性。抗癌药物开发成功率低的主要因素之一是临床前模型未能充分概括人类癌症的复杂性和异质性。出于通量和容量的原因，高通量筛选生长抑制测定几乎仅使用在含血清培养基中的组织培养物处理的塑料/玻璃表面上培养的肿瘤细胞系的二维（2D）单层。然而，这些2D肿瘤细胞系培养物未能重现实体瘤中细胞的三维（3D）背景，即使肿瘤微环境已显示对抗癌药物反应具有深远影响。肿瘤球体仍然是最好的特征和最广泛使用的3D模型;然而，球体大小往往是不均匀的，使它们不适合高通量药物测试。为了规避这一挑战，我们已经开发了使用非粘附性水凝胶的限定尺寸的微孔阵列，其适用于各种癌细胞系以制造尺寸受控的3D微肿瘤。我们证明，水凝胶微孔阵列平台可以成功应用于在3-6天内从许多宫颈和乳腺以及头颈部鳞状细胞癌（HNSCC）细胞中产生数百个均匀的微小肿瘤。此外，控制水凝胶阵列中微孔的尺寸允许精确控制微肿瘤的尺寸。最后，我们展示了该平台技术的应用，以探测激活以及抑制表皮生长因子受体（EGFR）信号在3D HNSCC微肿瘤中分别响应EGF和西妥昔单抗治疗。我们相信，使用水凝胶阵列产生大量均匀尺寸和3D形态的HNSCC微肿瘤的能力将提供更多的生理体外3D肿瘤模型，以研究肿瘤尺寸如何影响信号传导通路激活和癌症药物疗效。
Despite significant investments in cancer research and drug discovery/development, the rate of new cancer drug approval is ≤5% and most cases of metastatic cancer remain incurable. Ninety-five percent of new cancer drugs fail in clinical development because of a lack of therapeutic efficacy and/or unacceptable toxicity. One of the major factors responsible for the low success rate of anticancer drug development is the failure of preclinical models to adequately recapitulate the complexity and heterogeneity of human cancer. For throughput and capacity reasons, high-throughput screening growth inhibition assays almost exclusively use two-dimensional (2D) monolayers of tumor cell lines cultured on tissue culture-treated plastic/glass surfaces in serum-containing medium. However, these 2D tumor cell line cultures fail to recapitulate the three-dimensional (3D) context of cells in solid tumors even though the tumor microenvironment has been shown to have a profound effect on anticancer drug responses. Tumor spheroids remain the best characterized and most widely used 3D models; however, spheroid sizes tend to be nonuniform, making them unsuitable for high-throughput drug testing. To circumvent this challenge, we have developed defined size microwell arrays using nonadhesive hydrogels that are applicable to a wide variety of cancer cell lines to fabricate size-controlled 3D microtumors. We demonstrate that the hydrogel microwell array platform can be applied successfully to generate hundreds of uniform microtumors within 3–6 days from many cervical and breast, as well as head and neck squamous cell carcinoma (HNSCC) cells. Moreover, controlling size of the microwells in the hydrogel array allows precise control over the size of the microtumors. Finally, we demonstrate the application of this platform technology to probe activation as well as inhibition of epidermal growth factor receptor (EGFR) signaling in 3D HNSCC microtumors in response to EGF and cetuximab treatments, respectively. We believe that the ability to generate large numbers of HNSCC microtumors of uniform size and 3D morphology using hydrogel arrays will provide more physiological in vitro 3D tumor models to investigate how tumor size influences signaling pathway activation and cancer drug efficacy.
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影响因子：
11.2
作者：
Hielscher AC;Gerecht S
通讯作者：
Gerecht S
影响因子：
19
作者：
Yu, Hongmei;Mouw, Janna Kay;Weaver, Valerie M.
通讯作者：
Weaver, Valerie M.
影响因子：
6.1
作者：
Karp, Jeffrey M.;Yeh, Judy;Khademhosseini, Ali
通讯作者：
Khademhosseini, Ali
肿瘤血管生成的微环境调节：生物学和工程考虑
影响因子：
2.6
作者：
D. Infanger;S. Pathi;C. Fischbach
通讯作者：
C. Fischbach
球体抵抗光动力处理背后的机制：流式细胞术研究
影响因子：
3.3
作者：
C. West;J. Moore
通讯作者：
J. Moore