Stabilization of hESCs in two distinct substates along the continuum of pluripotency.

A detailed understanding of the developmental substates of human pluripotent stem cells (hPSCs) is needed to optimize their use in cell therapy and for modeling early development. Genetic instability and risk of tumorigenicity of primed hPSCs are well documented, but a systematic isogenic comparison between substates has not been performed. We derived four hESC lines in naive human stem cell medium (NHSM) and generated isogenic pairs of NHSM and primed cultures. Through phenotypic, transcriptomic, and methylation profiling, we identified changes that arose during the transition to a primed substate. Although early NHSM cultures displayed naive characteristics, including greater proliferation and clonogenic potential compared with primed cultures, they drifted toward a more primed-like substate over time, including accumulation of genetic abnormalities. Overall, we show that transcriptomic and epigenomic profiling can be used to place human pluripotent cultures along a developmental continuum and may inform their utility for clinical and research applications.

MSC-NTF (NurOwn®) exosomes: a novel therapeutic modality in the mouse LPS-induced ARDS model.

BACKGROUND: One of the most severe complications of the current COVID-19 pandemic is acute respiratory distress syndrome (ARDS). ARDS is caused by increased amounts of pro-inflammatory cytokines, leading to lung damage and loss of lung function. There are currently no effective therapies for combatting ARDS. Mesenchymal stem cells (MSCs) have been suggested as a potential treatment for ARDS due to their significant immunomodulatory properties. MSC small extracellular vesicles (sEVs), including exosomes, modulate the immune response as effectively as MSCs themselves, with the added advantages of increased safety and tissue penetration. METHODS: We isolated sEVs from MSCs induced to secrete increased levels of neurotrophic and immunomodulatory factors, termed Exo MSC-NTF, and compared their ability to treat ARDS, in a lung injury LPS mouse model, to sEVs isolated from naïve MSCs (Exo MSC). Measurments of lung histopathological changes and neutrophil infiltration, blood oxygen saturation, and bronchoalveolar lavge fluid (BALF) proinflammatory cytokines and coagulation related factors were performed. RESULTS: We found that Exo MSC-NTF was superior to Exo MSC in reducing LPS-induced ARDS markers, including physiological lung damage such as alveolar wall thickness, fibrin presence, and neutrophil accumulation, as well as increasing oxygenation levels. Furthermore, Exo MSC-NTF reversed the imbalance in the host immune response, seen as decreased IFN-γ, IL-6, TNF-α, and RANTES levels in the bronchoalveolar lavage fluid. CONCLUSIONS: These positive preclinical results suggest that Exo MSC-NTF may be suitable as a therapy for COVID-19-induced ARDS and are more effective at combatting ARDS physiological, pathological, and biochemical symptoms than sEVs isolated from non-induced MSCs.

Effect of an auditory feedback substitution, tactilo-kinesthetic, or visual feedback on kinematics of pouring water from kettle into cup.

Pouring hot water from a kettle into a cup may prove a hazardous task, especially for the elderly or the visually-impaired. Individuals with deteriorating eyesight may endanger their hands by performing this task with both hands, relaying on tactilo-kinesthetic feedback (TKF). Auditory feedback (AF) may allow them to perform the task singlehandedly, thereby reducing the risk for injury. However since relying on an AF is not intuitive and requires practice, we aimed to determine if AF supplied during the task of pouring water can be used naturally as visual feedback (VF) following practice. For this purpose, we quantified, in young healthy sighted subjects (n = 20), the performance and kinematics of pouring water in the presence of three isolated feedbacks: visual, tactilo-kinesthetic, or auditory. There were no significant differences between the weights of spilled water in the AF condition compared to the TKF condition in the first, fifth or thirteenth trials. The subjectively-reported difficulty levels of using the TKF and the AF were significantly reduced between the first and thirteenth trials for both TKF (p = 0.01) and AF (p = 0.001). Trunk rotation during the first trial using the TKF was significantly lower than the trunk rotation while using VF. Also, shoulder adduction during the first trial using the TKF was significantly higher than the shoulder adduction while using the VF. During the AF trials, the median travel distance of the tip of the kettle was significantly reduced in the first trials so that in the thirtieth trial it did not differ significantly from the median travel distance during the thirtieth trial using TKF and VF. The maximal velocity of the tip of the kettle was constant for each of the feedback conditions but was higher in 10 cm s(-1) using VF than TKF, which was higher in 10 cm s(-1) from using AF. The smoothness of movement of the TKF and AF conditions, expressed by the normalized jerk score (NJSM), was one and two orders of magnitude higher from the VF, respectively. The median NJSM then decreased significantly by the fifth trial. Monitoring in-house activity via motion capture and classification of movements, i.e. liquid pouring, can assist with daily activities via AF. As a built-in feature in a smart home, this task-specific AF may prevent burn injuries of the visually-impaired.