Abstract：
Microenvironmental mechanical signals are fundamental regulators of cell behavior both in physiological and in pathological context, particularly in the induction and maintenance of tumorigenic properties. It is thus of utmost importance to experimentally recreate conditions that mimic the physical attributes of real tissues to study their impact on cell behavior and in particular on the induction of cancer stem cell (CSC) properties. Here we present protocols to investigate the role of mechanical stiffness on reprogramming of primary mammary gland cells into CSCs, including the synthesis of hydrogel substrates of the desired stiffness, the isolation and culture of primary differentiated normal cells derived from the human mammary gland, and the assessment of their CSC attributes after oncogene-mediated transformation.
摘要：
微环境机械信号是生理和病理背景下细胞行为的基本调节因子。特别是在诱导和维持肿瘤特性。因此，最重要的是通过实验重建模拟真实组织的物理属性的条件，以研究它们对细胞行为的影响，特别是对癌症干细胞（CSC）特性的诱导。在这里，我们提出了研究机械刚度对原代乳腺细胞重编程为CSC的作用的方案，包括合成具有所需刚度的水凝胶基质，分离和培养来自人类乳腺的原代分化正常细胞，以及在癌基因介导的转化后对其CSC属性的评估。
