The success of human pluripotent stem cells (hPSCs) as a source of future cell therapies hinges, in part, on the availability of a robust and scalable culture system that can readily produce a clinically relevant number of cells and their derivatives. Stirred suspension culture has been identified as one such promising platform due to its ease of use, scalability, and widespread use in the pharmaceutical industry (e.g., CHO cell-based production of therapeutic proteins) among others. However, culture of undifferentiated hPSCs in stirred suspension is a relatively new development within the past several years, and little is known beyond empirically optimized culture parameters. In particular, detailed characterizations of different agitation rates and their influence on the propagation of hPSCs are often not reported in the literature. In the current study, we systematically investigated various agitation rates to characterize their impact on cell yield, viability, and the maintenance of pluripotency. Additionally, we closely examined the distribution of cell aggregates and how the observed culture outcomes are attributed to their size distribution. Overall, our results showed that moderate agitation maximized the propagation of hPSCs to approximately 38-fold over 7 days by keeping the cell aggregates below the critical size, beyond which the cells are impacted by the diffusion limit, while limiting cell death caused by excessive fluidic forces. Furthermore, we observed that fluidic agitation could regulate not only cell aggregation, but also expression of some key signaling proteins in hPSCs. This indicates a new possibility to guide stem cell fate determination by fluidic agitation in stirred suspension cultures. Biotechnol. Bioeng. 2017;114: 2109-2120. © 2017 Wiley Periodicals, Inc.
Introduction: Pneumothorax is an emergency having high mortality rate. Early recognition and management is crucial. Aim: To study the demographic data, clinical presentation, risk factor and management of cases of spontaneous pneumothorax presenting to ED. Material and Method: Observational analytical study done in 75 patients aged 20-74 years admitted at a tertiary care hospital from April 2018 to March 2019. Patients coming with spontaneous pneumothorx in ER were included in the study. History of trauma and iatrogenic causes were excluded.Result: Details about demographic data, clinical presentation, risk factor and management were recorded. Their main complaints were dyspnoea followed by chest pain. In our study out of 75 patients, ICD was inserted in 63 patients,7 patients managed with simple needle aspiration and 5 patients were treated conservatively. In our study 58 patients were shifted to E Ward/ICU, 8 patients were shifted to CT Sx ward, 5 patients shifted to ward, 3 patients took LAMA and 1 patient expired. Conclusion:In our study we found that SSP (secondary spontaneous pneumothorax) is more common than PSP (primary spontaneous pneumothorax), majority of patients were male and COPD (chronic obstructive pulmonary diseases) is the commonest cause of the SSP. Intercostal tube drainage is the main mode of management.
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