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开发一种微流控平台以维持培养中显微切割肿瘤切片的活力。

Development of a microfluidic platform to maintain viability of micro-dissected tumor slices in culture.

作者信息

Parsian Maryam, Mutlu Pelin, Yildirim Ender, Ildiz Can, Ozen Can, Gunduz Ufuk

机构信息

Department of Biotechnology, Middle East Technical University, Ankara, Turkey.

Department of Biotechnology, Ankara University, Ankara, Turkey.

出版信息

Biomicrofluidics. 2022 May 5;16(3):034103. doi: 10.1063/5.0087532. eCollection 2022 May.

Abstract

One of the issues limiting the development of personalized medicine is the absence of realistic models that reflect the nature and complexity of tumor tissues. We described a new tissue culture approach that combines a microfluidic chip with the microdissected breast cancer tumor. "Tumor-on-a-chip" devices are suitable for precision medicine since the viability of tissue samples is maintained during the culture period by continuously feeding fresh media and eliminating metabolic wastes from the tissue. However, the mass transport of oxygen, which arguably is the most critical nutrient, is rarely assessed. According to our results, transportation of oxygen provides satisfactory oxygenation within the system. A high level of dissolved oxygen, around 98%-100% for every 24 h, was measurable in the outlet medium. The microfluidic chip system developed within the scope of this study allows living and testing tumor tissues under laboratory conditions. In this study, tumors were generated in CD-1 mice using MDA-MB-231 and SKBR-3 cell lines. Microdissected tumor tissues were cultured both in the newly developed microfluidic chip system and in conventional 24-well culture plates. Two systems were compared for two different types of tumors. The confocal microscopy analyses, lactate dehydrogenase release, and glucose consumption values showed that the tissues in the microfluidic system remained more viable with respect to the conventional well plate culturing method, up to 96 h. The new culturing technique described here may be superior to conventional culturing techniques for developing new treatment strategies, such as testing chemotherapeutics on tumor samples from individual patients.

摘要

限制个性化医疗发展的问题之一是缺乏能够反映肿瘤组织性质和复杂性的现实模型。我们描述了一种将微流控芯片与显微切割的乳腺癌肿瘤相结合的新组织培养方法。“芯片上的肿瘤”装置适用于精准医疗,因为在培养期间,通过持续供应新鲜培养基并清除组织中的代谢废物,可维持组织样本的活力。然而,作为最关键营养素的氧气的质量传输却很少被评估。根据我们的结果,氧气传输在系统内提供了令人满意的氧合作用。在流出培养基中可测量到高水平的溶解氧,每24小时约为98%-100%。在本研究范围内开发的微流控芯片系统允许在实验室条件下培养和测试肿瘤组织。在本研究中,使用MDA-MB-231和SKBR-3细胞系在CD-1小鼠体内生成肿瘤。将显微切割的肿瘤组织在新开发的微流控芯片系统和传统的24孔培养板中培养。对两种不同类型的肿瘤比较了这两个系统。共聚焦显微镜分析、乳酸脱氢酶释放和葡萄糖消耗值表明,在长达96小时的时间里,相对于传统孔板培养方法,微流控系统中的组织保持了更高的活力。这里描述的新培养技术在开发新的治疗策略方面可能优于传统培养技术,例如对来自个体患者的肿瘤样本进行化疗药物测试。

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