Isolation of Primary Human Colon Tumor Cells from Surgical Tissues and Culturing Them Directly on Soft Elastic Substrates for Traction Cytometry

M. Yakut Ali; Sandeep V. Anand; Krishnarao Tangella; Davendra Ramkumar; Taher A. Saif

doi:10.3791/52532

JoVE Journal
Bioengineering

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JoVE Journal Bioengineering

Isolation of Primary Human Colon Tumor Cells from Surgical Tissues and Culturing Them Directly on Soft Elastic Substrates for Traction Cytometry

从手术组织小学的人结肠癌细胞的分离和培养直接它们柔软的弹性基底牵引术

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09:28 min

June 4, 2015

DOI: 10.3791/52532-v

M. Yakut Ali¹, Sandeep V. Anand¹, Krishnarao Tangella^2,3, Davendra Ramkumar^2,3, Taher A. Saif^1,4

¹Department of Mechanical Science and Engineering,University of Illinois at Urbana-Champaign, ²College of Medicine,University of Illinois at Urbana-Champaign, ³Provena Covenant Medical Centre, ⁴Micro and Nanotechnology Laboratory,University of Illinois at Urbana-Champaign

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描述了从外科结肠肿瘤和正常组织中提取原代人类细胞的方案。然后将分离的细胞培养在由细胞外基质蛋白功能化的软弹性底物（聚丙烯酰胺水凝胶）上，并用荧光微珠包埋。进行牵引细胞术以评估细胞收缩应激。

该程序的总体目标是从外科人结肠肿瘤以及正常组织中分离原代细胞，并将它们直接培养在具有不同弹性的基质上进行分析。这是通过首先从人结肠肿瘤和正常区域收集手术组织来实现的。接下来，碎组织的酶解离会产生许多单细胞。

最后一步是在包埋有纳米级荧光珠的软聚丙烯酰胺水凝胶和聚苯乙烯培养皿上培养分离的细胞。最后，进行牵引细胞术以获得空间细胞牵引图，并进行免疫荧光显微镜检查以可视化细胞骨架组织。该技术的主要优点是，从手术组织中分离的原代人结肠细胞可以直接在具有不同刚度的基质上培养，用于牵引细胞术和免疫荧光显微镜测定。

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生物工程 100期主人体结肠肿瘤细胞柔软的弹性基底牵引力显微镜力学生物学免疫荧光显微镜细胞力学