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Author:
Date:
2013-12-05
[Abstract] The use of three-dimensional (3D) cell culture systems is widely accepted as representing a more physiologically relevant means to propagate mammary epithelial and breast cancer cells. However, 3D cultures systems are plagued by several experimental and technical limitations as compared to their traditional 2D counterparts. For instance, quantifying the growth of mammary epithelial or breast cancer organoids longitudinally is particularly troublesome using standard [3H]thymidine or MTT assay systems, or using computer-assisted area calculations. Likewise, the nature of the ...
[摘要] 使用三维（3D）细胞培养系统被广泛接受为代表更多生理相关手段以增殖乳腺上皮和乳腺癌细胞。然而，与其传统的2D对应物相比，3D培养系统受到若干实验和技术限制的困扰。例如，使用标准的[3 H]胸苷或MTT测定系统或使用计算机辅助面积计算，纵向量化乳腺上皮或乳腺癌组织的生长是特别麻烦的。同样，在3D条件下由乳腺癌细胞形成的多细胞聚集体和类器体的性质阻止了细胞从3D基质的有效回收，这是耗时的并且导致假的结果的事件。这里描述的测定利用萤火虫荧光素酶的稳定表达作为定量在3D矩阵内传播的细胞的纵向生长的手段。该技术的主要优点包括其高通量性质和在规定的时间段内纵向跟踪单个孔的能力，从而降低与测定性能相关的成本。最后，该技术可以容易地与药物治疗和/或遗传操作组合以测定它们对3D类器官生长的影响。
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