oaioai:figshare.com:article/1369828

Adhesion of tumor cells to protein-immobilized microfluidic channels under low shear (0.35dyn/cm<sup>2</sup>).

Abstract

<p>Microfluidic channels were incubated with Protein G (100μg/ml), then anti-TF IgG (100μg/ml), or an anti-His antibody (100μg/ml) followed by TFPI (100μg/ml). Isotype IgG (100μg/ml) and anti-His IgG (100μg/ml) antibodies were used as negative control for anti-TF IgG and TFPI respectively. Tumor cells (1x10<sup>6</sup>cells/mL, pre-treated with 10nM FVIIa and 10nM FX for TFPI-coated channels) were introduced into the channels at 0.35dyn/cm<sup>2</sup> for 30 minutes, and non-specifically adhered cells were removed at 2.0dyn/cm<sup>2</sup>. The entire channel was imaged to quantify the number of adherent cells. <b>A.</b> Representative bright field images of adherent tumor cells on channels immobilized with Protein G (negative control), anti-TF IgG and TFPI showing that more MDA-MB-231 than MCF-7 cells were bound to both anti-TF IgG- and TFPI-coated channels. <b>B.</b> The number of adherent cells was counted and normalized by the channel area. MDA-MB-231 showed significantly higher adhesion to TFPI- and anti-TF IgG-coated channels than MCF-7 (* p < 0.05, n = 4 for anti-TF IgG, n = 3 for TFPI). Significantly more MDA-MB-231 bound to TFPI- and anti-TF IgG-coated channels than negative controls (** p<0.05). <b>C.</b> MDA-MB-231 cells were pretreated with 50μg/ml anti-TF IgG (TF9-5B7 which blocks FVIIa binding to TF, or TF9-10H10 which does not block FVIIa binding to TF). The positive control had no antibody pretreatment, and isotype IgG pretreatment (50μg/ml) was used as a negative control. Blocking FVIIa binding to TF with TF9-5B7 antibody significantly decreased adhesion to TFPI-coated channels (* p < 0.05, n = 4). The observed decrease in MDA-MB-231 adhesion with the TF9-10H10 antibody, albeit not significant with this stringent statistical test, could be due to steric hindrance of TFPI binding to the TF/FVIIa/FXa complex on the tumor cells.</p

Similar works

Full text

thumbnail-image
oaioai:figshare.com:article/1369828Last time updated on 2/12/2018

This paper was published in FigShare.

Having an issue?

Is data on this page outdated, violates copyrights or anything else? Report the problem now and we will take corresponding actions after reviewing your request.