Simple and cost-effective assay for isolating invasive living cells

BioTechniques
Takahisa TakinoMotoharu Seiki

Abstract

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.

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Citations

Feb 6, 2020·Current Protocols in Cell Biology·Takahisa TakinoMotoharu Seiki
Oct 21, 2019·Biochemical and Biophysical Research Communications·Hirari YamahanaYoshihiro Uto
Aug 13, 2021·Biochemistry and Biophysics Reports·Hirari YamahanaTakahisa Takino

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Methods Mentioned

BETA
PCR
Fluorescence
electrophoresis
zymography
confocal microscopy

Software Mentioned

Image J
ImageJ

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