Colorectal tumor 3D in vitro models: advantages of biofabrication for the recapitulation of early stages of tumour development
Colorectal tumor 3D in vitro models: advantages of biofabrication for the recapitulation of early stages of tumour development
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DOI：
10.1088/2057-1976/aac1c9
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发表时间：
2018-07-01
影响因子：
1.4
通讯作者：
Tirella, A.
中科院分区：
文献类型：
--
作者：
de la Rosa, J. M. Rios;Wubetu, J.;Tirella, A.
大多数癌症相关的体外研究是在细胞单层或球状体上进行的。虽然这种方法已经导致了关键的发现，但它在概括肿瘤发展的不同阶段方面仍然效果不佳。新型三维（3D）系统及其制造技术方法的出现将改善该领域，为肿瘤发展和治疗的研究提供更具生理相关性和高通量的体外系统。在这里，我们描述了基于藻酸盐的3D模型的制作，该模型概括了结直肠癌的早期阶段，跟踪肿瘤发展的两个主要生物标志物：CD 44和HIF-1 α。我们优化了制造过程，以获得具有可控尺寸和刚度的藻酸盐微珠，模拟结直肠癌的早期阶段。将人结肠直肠HCT-116癌细胞以受控的初始数量包封，并将所述3D体外模型的细胞活力和蛋白质表达与当前金标准（细胞单层和球状体）的细胞活力和蛋白质表达进行比较。我们的结果证明，与球状体培养物相比，包封的HCT-116表现出高活力、干细胞样细胞群的增加（CD 44表达增加）和缺氧区域的减少（HIF-1 α表达降低）。总之，我们表明，我们的生物制造系统是一种高度可重复且易于获得的替代方法来研究细胞行为，与其他体外模型相比，可以更好地模拟结直肠癌的早期阶段。使用生物制造的体外模型将提高结果的可翻译性，特别是在测试治疗干预策略时。
The majority of cancer-related in vitro studies are conducted on cell monolayers or spheroids. Although this approach has led to key discoveries, it still has a poor outcome in recapitulating the different stages of tumor development. The advent of novel three-dimensional (3D) systems and technological methods for their fabrication is set to improve the field, offering a more physiologically relevant and high throughput in vitro system for the study of tumor development and treatment. Here we describe the fabrication of alginate-based 3D models that recapitulate the early stages of colorectal cancer, tracking two of the main biomarkers for tumor development: CD44 and HIF-1 alpha. We optimized the fabrication process to obtain alginate micro-beads with controlled size and stiffness, mimicking the early stages of colorectal cancer. Human colorectal HCT-116 cancer cells were encapsulated with controlled initial number, and cell viability and protein expression of said 3D in vitro models was compared to that of current gold standards (cell monolayers and spheroids). Our results evidenced that encapsulated HCT-116 demonstrated a high viability, increase in stem-like cell populations (increased expression of CD44) and reduced hypoxic regions (lower HIF-1 alpha expression) compared to spheroid cultures. In conclusion we show that our biofabricated system is a highly reproducible and easily accessible alternative to study cell behavior, allowing to better mimic the early stages of colorectal cancer in comparison to other in vitro models. The use of biofabricated in vitro models will improve the translatability of results, in particular when testing strategies for therapeutic intervention.