Real-Time Cell Invasion Assay

Cell invasion assay was assessed using the xCELLigence Real Time Cell Analysis (RTCA) technology (Acea Bioscience) and E-16-well plates (#05469830001) as described [35 (link)]. Bottom wells were coated with 20 μg/well matrigel (Corning^® #356231) diluted in serum-free medium. Matrigel was allowed to polymerize for 1 h at 37 °C prior to seed MLPS cells (1 × 10⁴ cells/well) suspended in CM from monocytes pre-cultured with MLPS cells or CM from monocytes alone, the last employed as a control. In both cases, CMs were supplemented with 10% heat-inactivated human serum. Cells that cross matrigel adhere to the bottom of plates causing impedance changes which are proportional to the number of invading cells. The impedance value of each well was automatically monitored in real-time for 18 h and expressed as a cell index value.

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Minopoli M., Sarno S., Cannella L., Tafuto S., Scognamiglio G., Gallo M., Fazioli F., Azzaro R., Apice G., De Angelis B., Tamborini E., Garofalo C., Pignochino Y., Mercatali L., Ibrahim T., Falcioni R., Valenti B., Maestro R., Scotlandi K., De Chiara A, & Carriero M.V. (2021). Crosstalk between Macrophages and Myxoid Liposarcoma Cells Increases Spreading and Invasiveness of Tumor Cells. Cancers, 13(13), 3298.

Publication 2021

xCELLigence Real Time Cell Analysis (RTCA) technology E-16-well plates

Assay Cell Cultured cells Human Matrigel Monocytes Serum

Corresponding Organization :

Other organizations : Istituto Nazionale Tumori IRCCS "Fondazione G. Pascale", Bambino Gesù Children's Hospital, Fondazione IRCCS Istituto Nazionale dei Tumori, Istituto Oncologico Veneto, Istituti di Ricovero e Cura a Carattere Scientifico, University of Turin, Istituto Scientifico Romagnolo per lo Studio e la Cura dei Tumori, Centro di Riferimento Oncologico, Istituto Ortopedico Rizzoli

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Variable analysis

independent variables

Conditioned medium (CM) from monocytes pre-cultured with MLPS cells
CM from monocytes alone (control)

dependent variables

Cell invasion across matrigel, measured as impedance changes and expressed as cell index value

control variables

Matrigel coating (20 μg/well) in serum-free medium, allowed to polymerize for 1 h at 37 °C prior to seeding cells
MLPS cells seeded at 1 × 10^4 cells/well
CMs supplemented with 10% heat-inactivated human serum

controls

Positive control: CM from monocytes pre-cultured with MLPS cells
Negative control: CM from monocytes alone

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