Three-dimensional Invasion Assay
三维侵袭实验
发布: 2013年09月05日第3卷第17期 DOI: 10.21769/BioProtoc.885 浏览次数: 12485
评审: Lin FangFanglian HeAnonymous reviewer(s)
PDF 
Q&A
参见作者原研究论文
The authors used this protocol in:
Apr 2012
Abstract
The invasive ability of cancer cells is a crucial function for cancer metastasis and the surrounding microenvironment of cancer cells in living tissues is three-dimension (3D). Therefore, to establish an in vitro invasion assay in a 3D system to predict cancer invasive ability is valuable in the research for cancer metastasis. Here, we describe a 3D invasion assay for observing the morphology and comparing the invasive ability of cancer cells in artificial 3D environments (Yang et al., 2012). Collagen I gels are used to cover on the top of cancer cells attached on coverslip glass dish and medium containing FBS is added as a chemoattractant. After incubation for a suitable time, the cells are fixed and stained. The invasion index can be calculated and the morphology can be imaged with a laser confocal microscope.
Collagen gel (胶原凝胶)
Invasion index (侵袭指数)
Materials and Reagents
- Cell lines: OECM-1(Huang et al., 2004) and FaDu (ATCC® HTB-43 TM)
- 0.1% Trypsin-EDTA (Life Technologies, Gibco®, catalog number: 15400 )
- 0.1 mg/ml poly-L lysine (Sigma-Aldrich, catalog number: P9404-25MG )
- PBS
- FBS (Thermo Fisher Scientific, catalog number: SH30071.03 )
- PureCor® bovine collagen solution (Advance Biomatrix Inc., catalog number: 5005-B )
- 1 M NaOH solution
- 5x RPMI medium
- 3% Paraformaldehyde (Sigma-Aldrich, catalog number: P6148-500G )
- 0.5% Triton X100 (Bionovas, catalog number: 56-81-5 )
- Alexa Fluor® 488 Phalloidin (Life Technologies, catalog number: A12379 )
- DAPI (Sigma-Aldrich, catalog number: D8417 )
- 1% BSA in PBS
- 1.8 mg/ml collagen I mix solution (see Recipes)
Equipment
- Lab-Tek® chambered #1.0 coverglass system (NUNC, catalog number: 155383 )
- Laser confocal microscope with 60x oil lens (Olympus, model: FV1000 )
- CO2 incubator
Software
- Olympus FV10-ASW 1.7 software
Procedure
文章信息
版权信息
© 2013 The Authors; exclusive licensee Bio-protocol LLC.
如何引用
Yang, W. and Yang, M. (2013). Three-dimensional Invasion Assay. Bio-protocol 3(17): e885. DOI: 10.21769/BioProtoc.885.
Download Citation in RIS Format
分类
癌症生物学 > 通用技术 > 肿瘤微环境
癌症生物学 > 侵袭和转移 > 细胞生物学试验
细胞生物学 > 细胞成像 > 共聚焦显微镜
您对这篇实验方法有问题吗？
在此处发布您的问题，我们将邀请本文作者来回答。同时，我们会将您的问题发布到Bio-protocol Exchange，以便寻求社区成员的帮助。
提问指南
+ 问题描述
写下详细的问题描述，包括所有有助于他人回答您问题的信息（例如实验过程、条件和相关图像等）。
发布问题
