Tryple express cell dissociation reagent

All cell lines used for characterization were grown to 30% to 60% confluence at 37 °C in a humid atmosphere of 5% CO 2 . Cell lines were evaluated for mycoplasma contamination by PCR (Venor GeM Mycoplasma Detection Kit) and authenticated by IDEXX BioResearch. MCF7 (breast carcinoma, ATCC) cells were cultured in RPMI medium supplemented with 10% fetal bovine serum (FBS) previously inactivated at 56 °C for 30 min, 1% penicillin/streptomycin (Invitrogen), and 0.01 mg/mL human recombinant insulin (Life Technologies). MDA-MB-231 (breast carcinoma, ATCC) cells were grown in DMEM medium supplemented with 10% inactivated FBS and 1% penicillin/streptomycin. HCC827 (lung carcinoma, ATCC) cells and H1975 (lung carcinoma, ATCC) cells were grown in RPMI medium supplemented with 10% inactivated FBS and 1% penicillin/ streptomycin. Adherent cells were dissociated with TrypLE express cell dissociation reagent (Gibco) and resuspended in complete media. Their viability and average diameter were measured using acridine orange, propidium iodide solution on a Cellometer K2 (Nexcelom). Cells (from 50 to 500 cells depending on the experiment) were then spiked into 0.5 to 4.0 mL healthy human blood diluted 10× in PBS pH 7.2, prior to injection through the chip.