Figure 7.
In vitro aggregation of platelet from NSML patients and ex vivo thrombus formation of blood from NSML patients or treated with SHP2 inhibitor. (A) Platelets from healthy controls and NSML patients were stimulated with collagen (0.75 and 3.3 µg/mL). Platelet aggregation curves of a representative healthy control and from each of the 3 NSML patients are shown. Graph represents the percentage of maximal aggregation of 3 healthy controls and 3 NSML patients. Each point corresponds to an individual. Patient 1 is represented in green, patient 2 in red, and patient 3 in blue. (B) Anticoagulated human whole blood, where platelets were labeled with DiOC6, was perfused through Cellix Vena8 Fluoro+ biochips at different shear rates on fibrillar collagen matrix over 3 minutes. Thrombus formation was visualized in real time by videomicroscopy. Shown are representative 2- and 3-dimensional images of NSML patient 2 blood perfusion at a stenosis shear rate of 3000 s−1 obtained after processing with ImageJ and Imaris software, respectively (scale bar, 20 μm). Surface covered by platelets and thrombus volume were quantified at different shear rates (500, 1500, and 3000 s−1) by using ImageJ software. Graphs represent mean ± SEM for 3 healthy controls and 3 NSML patients. For NSML patients, each point corresponds to an individual (patient 1 is represented in green, patient 2 in red, and patient 3 in blue). (C) Anticoagulated human whole blood treated with SHP099 (10 μM) or its vehicle, dimethyl sulfoxide, for 2 hours and labeled with DiOC6 was perfused through Cellix Vena8 Fluoro+ biochips at 1500s−1 on fibrillar collagen matrix over 3 minutes. Thrombus formation was visualized in real time by videomicroscopy. Shown are representative 2- and 3-dimensional images of blood from 7 healthy donors. Surface covered by platelets and thrombus volume were quantified by using ImageJ software. Graphs represent mean ± SEM for 7 healthy donors. *P < .05, **P < .01, ***P < .001 vs healthy controls according to 2-way ANOVA and 1-sample t test. †P < .001 vs vehicle according to 1-sample t test.

In vitro aggregation of platelet from NSML patients and ex vivo thrombus formation of blood from NSML patients or treated with SHP2 inhibitor. (A) Platelets from healthy controls and NSML patients were stimulated with collagen (0.75 and 3.3 µg/mL). Platelet aggregation curves of a representative healthy control and from each of the 3 NSML patients are shown. Graph represents the percentage of maximal aggregation of 3 healthy controls and 3 NSML patients. Each point corresponds to an individual. Patient 1 is represented in green, patient 2 in red, and patient 3 in blue. (B) Anticoagulated human whole blood, where platelets were labeled with DiOC6, was perfused through Cellix Vena8 Fluoro+ biochips at different shear rates on fibrillar collagen matrix over 3 minutes. Thrombus formation was visualized in real time by videomicroscopy. Shown are representative 2- and 3-dimensional images of NSML patient 2 blood perfusion at a stenosis shear rate of 3000 s−1 obtained after processing with ImageJ and Imaris software, respectively (scale bar, 20 μm). Surface covered by platelets and thrombus volume were quantified at different shear rates (500, 1500, and 3000 s−1) by using ImageJ software. Graphs represent mean ± SEM for 3 healthy controls and 3 NSML patients. For NSML patients, each point corresponds to an individual (patient 1 is represented in green, patient 2 in red, and patient 3 in blue). (C) Anticoagulated human whole blood treated with SHP099 (10 μM) or its vehicle, dimethyl sulfoxide, for 2 hours and labeled with DiOC6 was perfused through Cellix Vena8 Fluoro+ biochips at 1500s−1 on fibrillar collagen matrix over 3 minutes. Thrombus formation was visualized in real time by videomicroscopy. Shown are representative 2- and 3-dimensional images of blood from 7 healthy donors. Surface covered by platelets and thrombus volume were quantified by using ImageJ software. Graphs represent mean ± SEM for 7 healthy donors. *P < .05, **P < .01, ***P < .001 vs healthy controls according to 2-way ANOVA and 1-sample t test. †P < .001 vs vehicle according to 1-sample t test.

Close Modal
This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal