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

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)

1. Open a new dual-chamber cell analyzer plate. Set aside the top chamber with electrodes.
2. Using a milling machine, shave off 2 mm of the U-shaped bottom wells of the cell analyzer plate.
3. 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.
4. Using a milling machine, cut out two longitudinal slits (1.5 mm x 5.6 mm) along the sides to snap into the ridges of the newly fabricated third chamber.
5. Using a milling machine, create a t....

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 ch.......

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
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