Three-Chambered Array-Based Impedance Assay: A Real-Time Analysis Technique to Assess Invasive Potential of Cancer Cells by Measuring Electrical Impedance

Published: April 30, 2023

Abstract

Source: Sharif, G. M. et al., Real-Time Detection and Capture of Invasive Cell Subpopulations from Co-Cultures. J. Vis. Exp. (2022).

This video describes an invasion assay using a three-chambered array based on electrical impedance. This technique measures the invasive potential of cancer cells under the influence of soluble factors secreted by the resident stromal cells in tumor microenvironments.

Protocol

1. New chamber design (Figure 1) Open a new dual-chamber cell analyzer plate. Set aside the top chamber with electrodes. Using a milling machine, shave off 2 mm of the U-shaped bottom wells of the cell analyzer plate. Attach a 2 cm x 7 cm polyethersulfone (PES) membrane with 0.2 μm pore size to the bottom of the shaved wells using UV-curated adhesive. Allow 30 min curation time to ensure the glue is completely cured and inert. Using a milling machine, cut out two longitudina…

Representative Results

Figure 1: Images of the array chambers and modifications. (A) The three chambers used to build the array. No modification was made on the top chamber harboring the electrodes. (B) From the middle chamber wells, a height of 2 mm has been shaved off and a membrane attached to the open bottom; longitudinal slits (1.5 mm x 5.6 mm) were add…

Declarações

The authors have nothing to disclose.

Materials

0.05% Trypsin-EDTA Thermofisher 25300-054
Adhesive Norland Optical Adhesive  NOA63
Bovine serum albumin (BSA) Sigma A9418
Cell lifter Sarstedt 83.1832
Cholera Toxin from Vibrio cholerae Thermofisher  12585-014
CIM-plate Agilent 5665817001 Cell analyzer plate
Collagenase from Clostridium histolyticum Sigma C0130
DMEM Thermofisher  11995-065
DMEM-F12 Thermofisher 11875-093
Fetal Bovine Serum (FBS), Heat Inactivated Omega Scientific  FB-12
HEPES  Thermofisher 15630106
Horse serum (HS) Gibco 16050-122
Human EGF Peprotech AF-100-15
Hydrocortisone  Sigma  H4001
Insulin Transferrin Selenium Ethanolamine (ITSX) (100x) Thermofisher 51500056
Insulin, Human Recombinant, Zinc Solution Sigma C8052
J2 Fibroblasts Stemcell (RRID:CVCL_W667)  100-0353
LymphoPrep Stemcell 7851 Density gradient medium for the isolation of mononuclear cells
Matrigel Corning 354230 Basement membrane matrix
MCFDCIS.com cells ( DCIS) RRID:CVCL_5552
MDA-MB-231 cells  RRID:CVCL_0062
Milling machine  Bridgeport Series 1 Vertical
Phosphate-buffered saline (1x) Thermofisher 10010049
Polyethersulfone (PES) membrane Sterlitech PCTF029030
RBC lysis solution Stemcell 7800
RTCA DP analyzer Agilent 3X16  Dual purpose cell analyzer
Trypsin Sigma  T4799

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Three-Chambered Array-Based Impedance Assay: A Real-Time Analysis Technique to Assess Invasive Potential of Cancer Cells by Measuring Electrical Impedance. J. Vis. Exp. (Pending Publication), e21156, doi: (2023).

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