Simple and cost-effective assay for isolating invasive living cells

BioTechniques

Hiroshi Sato¹  Takahisa Takino²  Motoharu Seiki³ 

[1] Science, Kanazawa University, Kakuma-machi, Kanazawa 920-1192, Japan;;1Division of Education for Global Standard, Institute of Liberal Arts &;2Division of Molecular Virology &

关键词: collagen gel;    invasion assay;    matrix metalloproteinase;    three dimensional;   

DOI  :  10.2144/btn-2018-0036

来源: DOAJ

【 摘 要 】

A modified invasion assay using a three-dimensional collagen gel was developed that enables isolation of invasive living cells; it was named the invading cell trapping (iCT) assay. A small cell strainer consisting of a nylon mesh with 40-μm2 pores was used, and collagen gel layers formed across the membrane. Test cells were seeded in the lower gel layer and invasive cells were attracted upward and trapped in the upper gel. After incubation, the collagen gel layers in cell strainers were easily separated and living cells in the gel were counted and analyzed. An advantage of the iCT assay is that it can capture living invasive cells in the upper gel while leaving noninvasive ones in the lower layer. Further enrichment of the two cell populations can be achieved by repeating the assay. Thus, the iCT assay allows comparative analysis of invasive versus noninvasive cells.

【 授权许可】

Unknown