Functional Interrogation of Ca2+ Signals in Human Cancer Cells In Vitro and Ex Vivo by Fluorescent Microscopy and Molecular Tools.
Functional Interrogation of Ca2+ Signals in Human Cancer Cells In Vitro and Ex Vivo by Fluorescent Microscopy and Molecular Tools.
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通过荧光显微镜和分子工具对体外和离体人类癌细胞中的 Ca2 信号进行功能询问。
DOI：
10.1007/978-1-0716-3271-0_7
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发表时间：
2023
期刊：
影响因子：
--
通讯作者：
Tang,Xin
中科院分区：
文献类型：
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作者：
Liang,Chenyu;Huang,Miao;Tanaka,Mai;Lightsey,Suzanne;Temples,Madison;Lepler,SharonE;Sheng,Peike;Mann,WilliamP;Widener,AdrienneE;Siemann,DietmarW;Sharma,Blanka;Xie,Mingyi;Dai,Yao;Phelps,Edward;Zeng,Bo;Tang,Xin
遗传编码钙指示剂（GECIs）和高分辨率共聚焦显微镜使细胞和组织中钙信号的动态可视化成为可能。二维和三维生物相容性材料以可编程的方式模拟肿瘤和健康组织的机械微环境。癌症异种移植模型和肿瘤切片的离体功能成像揭示了不同进展阶段肿瘤中钙动力学的生理相关功能。这些强大技术的整合使我们能够量化，诊断，建模和理解癌症病理生物学。在这里，我们描述了用于建立这种集成询问平台的详细材料和方法，从产生稳定表达CaViar（GCaMP 5G + QuasAr 2）的转导癌细胞系到2D/3D水凝胶和肿瘤组织中细胞的体外和离体钙成像。这些工具为详细探索生命系统中的机械-电化学网络动力学提供了可能性。
Genetically encoded calcium indicators (GECIs) and high-resolution confocal microscopy enable dynamic visualization of calcium signals in cells and tissues. Two-dimensional and 3D biocompatible materials mimic the mechanical microenvironments of tumor and healthy tissues in a programmable manner. Cancer xenograft models and ex vivo functional imaging of tumor slices reveal physiologically relevant functions of calcium dynamics in tumors at different progression stages. Integration of these powerful techniques allows us to quantify, diagnose, model, and understand cancer pathobiology. Here, we describe detailed materials and methods used to establish this integrated interrogation platform, from generating transduced cancer cell lines that stably express CaViar (GCaMP5G + QuasAr2) to in vitro and ex vivo calcium imaging of the cells in 2D/3D hydrogels and tumor tissues. These tools open the possibility for detailed explorations of mechano-electro-chemical network dynamics in living systems.