axion maestro edge  (Axion BioSystems)

Journal: bioRxiv

Article Title: Pseudomonas aeruginosa- mediated cardiac dysfunction is driven by extracellular vesicles released during infection

doi: 10.1101/2024.11.22.624948

Figure Lengend Snippet: hiPSC-CMs were plated in 24 well MEA plates, and the cells were exposed to the mixture of cardiomyocyte culture medium and C-media (1:1 ratio) harvested from uninfected (UI C-media) or P.a. infected ( P.a. C-media) hMDMs. The cardiomyocyte contractility and electrical activity were recorded using AxIS Navigator on the MEA system at 5% CO2 and at 37°C for indicated time points. Data analysis was performed using the cardiac analysis tool. ( A ) Electrical activity map showing the changes in the beat rate of cardiomyocytes treated with UI C-media and P.a. C-media. The activity map shown is a representative well from quadruplicate samples for each treatment and four repeats (N=4). ( B ) Representative traces recorded with the MEA system showing the beat period, T-wave, and FPD in UI-C-media and P.a. C-media-treated hiPSC-CMs. The graphs shown in ( C ) are beats per minute and ( D ) is filed potential duration (FPD) at baseline, 30-,45-, 60- and 120 minutes post treatment of hiPSC-CMs with UI C-media and P.a. C- media. ( E) Representative wave propagation of hiPSC-CMs treated with UI C-media and P.a. C-media. ( F-G) Representative EKG traces of hiPSC-CMs treated with UI C-media ( H-I ) P.a. C-media. Data shown in Figures C and D are accumulative data from 4 independent experiments, mean ± SD: * p<0.05, ** p <0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: In brief, cells were stabilized for a minimum of 30 min in the MEA system (Maestro Edge, Axion Biosystem, GA, USA), then the baseline was recorded, followed by C-media treatment.

Techniques: Infection, Activity Assay

Journal: bioRxiv

Article Title: Pseudomonas aeruginosa- mediated cardiac dysfunction is driven by extracellular vesicles released during infection

doi: 10.1101/2024.11.22.624948

Figure Lengend Snippet: hiPSC-CMs were plated in 24 well MEA plates, and the cells were exposed to the mixture of cardiomyocyte culture medium and C-media (1:1 ratio) harvested from hMDMs that were left uninfected (UI C-media) or P.a. infected (P.a. C-media) and heat-inactivated C-media (at 65°C for 45 min). The cardiomyocyte contractility and electrical activity was recorded using AxIS Navigator on MEA system at 5% CO2 and 37°C for indicated time points. Data analysis was performed using the cardiac analysis tool. ( A ) Electrical activity map showing the changes in the beat rate., Shown is a representative well from quadruplicate wells from each treatment (N=3). ( B ) A typical Cardiomyocyte beat on MEA. ( C ) Representative cardiomyocyte beat overlay recorded with the MEA system showing the beat period, T-wave, and FPD in UI C-media, P.a. C-media, and heat-inactivated C-media treated hiPSC-CMs. The graphs shown in ( D ) are beat rate, and ( E ) are field potential duration (FPD) at baseline, 30-, 60-, and 120 minutes post-treatment of hiPSC-CMs. Data shown in figures D and E are accumulative data from 3 independent experiments. mean ± SD; * p<0.05, ** p <0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: In brief, cells were stabilized for a minimum of 30 min in the MEA system (Maestro Edge, Axion Biosystem, GA, USA), then the baseline was recorded, followed by C-media treatment.

Techniques: Infection, Activity Assay

Journal: bioRxiv

Article Title: Pseudomonas aeruginosa- mediated cardiac dysfunction is driven by extracellular vesicles released during infection

doi: 10.1101/2024.11.22.624948

Figure Lengend Snippet: hiPSC-CMs were plated in 24 well MEA plates, and the cells exposed to the mixture of cardiomyocyte culture medium and C-media (1:1 ratio) harvested from uninfected (UI C-media), P.a. infected ( P.a. C-media) or heat-killed P.a. infected (HK P.a . C-media) hMDMs. The cardiomyocyte contractility and electrical activity was recorded using AxIS Navigator on the MEA system at 5% CO2 and at 37°C for indicated time points. Data analysis was performed using the cardiac analysis tool. ( A ) Electrical activity map showing the changes in beat rate of cardiomyocytes treated with UI C-media and P.a. C- media. The activity map shown is a representative well from quadruplicate samples for each treatment and three repeats (N=3). ( B ) Representative traces recorded with the MEA system showing the beat period, T-wave, and FPD in UI-C-media, P.a. C-media, and HK P.a. C-media-treated hiPSC-CMs. The graph shown in ( C ) is beats per minute, and ( D ) filed potential duration (FPD) at baseline, 30-, 45-, 60- and 120 min post-treatment of hiPSC-CMs with UI C-media, P.a. C-media, and HK P.a. C-media. Graphs shown in are ( E ) IL-1β, ( F) IL-6, ( G) TNF-α, and ( H) IL-10 levels in C-media harvested from uninfected, live P.a. and heat-killed P.a. infected hMDMs. Data shown in Figures C and D are accumulative data from 3 independent experiments (mean ± SD; ** p <0.01, *** p < 0.001, **** p < 0.0001). Data shown in figure E-H is representative data from two independent experiments, mean ± SD: * p<0.05, ** p <0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: In brief, cells were stabilized for a minimum of 30 min in the MEA system (Maestro Edge, Axion Biosystem, GA, USA), then the baseline was recorded, followed by C-media treatment.

Techniques: Infection, Activity Assay

Journal: bioRxiv

Article Title: Pseudomonas aeruginosa- mediated cardiac dysfunction is driven by extracellular vesicles released during infection

doi: 10.1101/2024.11.22.624948

Figure Lengend Snippet: hiPSC-CMs were plated in 24 well MEA plates, and the cells were exposed to P.a. (1 MOI) or left uninfected (UI). The cardiomyocyte contractility and electrical activity was recorded using AxIS Navigator on the MEA system at 5% CO2 and at 37°C for indicated time points. Data analysis was performed using the cardiac analysis tool. ( A ) Electrical activity map showing the changes in the beat rate of cardiomyocytes infected with P.a. or control (UI). The activity map shown is a representative well from quadruplicate samples from each treatment (N=3). ( B ) A representative cardiomyocyte beat overlay was recorded with the MEA system, showing the beat period, T-wave, and FPD in the UI and P.a. infected hiPSC-CMs. The graphs shown in ( C ) are beat rate, and ( D ) are field potential duration (FPD) at baseline, 5h, 6h, 7h, and 8h post-infection of hiPSC-CMs with P.a . or UI. Data shown in figures D, E, and F are accumulative data from 3 independent experiments; mean ± SD; * p<0.05, ** p <0.01, *** p < 0.001, **** p < 0.001, *** p < 0.0001. ( G ) Representative Phase-contrast images were captured at 8h post- infection with P.a. or left uninfected. Scalebar=50µm. The images shown are representative of three independent experiments.

Article Snippet: In brief, cells were stabilized for a minimum of 30 min in the MEA system (Maestro Edge, Axion Biosystem, GA, USA), then the baseline was recorded, followed by C-media treatment.

Techniques: Activity Assay, Infection, Control

Journal: bioRxiv

Article Title: Pseudomonas aeruginosa- mediated cardiac dysfunction is driven by extracellular vesicles released during infection

doi: 10.1101/2024.11.22.624948

Figure Lengend Snippet: hiPSC-CMs were cultured on the MEA plate and exposed with OMVs or left untreated (UT), and the cardiomyocyte contractility and electrical activity were recorded using AxIS Navigator on the MEA system at 5% CO2 and at 37°C for indicated time points. ( A ) Representative traces recorded with the MEA system showing the beat period, T- wave, and FPD in UT and OMV-treated hiPSC-CMs. The graphs shown in ( B ) are beats per minute and ( C ) FPD at baseline, 15-, 30-, 45-, and 60 min post-treatment of hiPSC-CMs with OMVs. ( D-E ) Representative beat period of untreated ( F-G ) OMV treated hiPSC- CMs over time. Data shown in Figures B and C are representative of 4 independent experiments, mean ± SD: * p<0.05, ** p <0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: In brief, cells were stabilized for a minimum of 30 min in the MEA system (Maestro Edge, Axion Biosystem, GA, USA), then the baseline was recorded, followed by C-media treatment.

Techniques: Cell Culture, Activity Assay

Journal: bioRxiv

Article Title: Pseudomonas aeruginosa- mediated cardiac dysfunction is driven by extracellular vesicles released during infection

doi: 10.1101/2024.11.22.624948

Figure Lengend Snippet: P.a. C-medium was collected as described earlier. For culturing P.a. without hMDMs, P.a. was grown in RPMI supplemented with 10% autologous serum (serum used for hMDM culture) for 2 days, and C-media was processed as described earlier. hiPSC-CMs were plated in 12 well-sterile plates, and the cells were exposed to the mixture of cardiomyocyte culture medium and C-media (1:1 ratio) harvested from uninfected (UI C-media), P.a. infected ( P.a. C-media) hMDMs or P.a RPMI C-media without hMDMs. The cardiomyocyte contractility and electrical activity were recorded using AxIS Navigator on the MEA system at 5% CO2 and at 37°C for indicated time points. Data analysis was performed using the cardiac analysis tool. ( A ) Electrical activity map showing the changes in the beat rate of cardiomyocytes infected with P.a. C-media with/without hMDMs or control (UI). The activity map shown is a representative well from quadruplicate samples from each treatment (N=3). ( B ) A representative cardiomyocyte beat overlay was recorded with the MEA system, showing the beat period, T-wave, and FPD from different experimental conditions as described above.

Article Snippet: In brief, cells were stabilized for a minimum of 30 min in the MEA system (Maestro Edge, Axion Biosystem, GA, USA), then the baseline was recorded, followed by C-media treatment.

Techniques: Sterility, Infection, Activity Assay, Control