Low Cost, Adhesion Strength Based Cell Sorter

In many in vitro experiments, primary cells are harvested from an animal species to undergo experimental manipulation and subsequent analysis. Cell sorters are a luxury to have after cell harvesting to ensure pure populations of cells. Recently, it was discovered that different cell types adhere to cell culture dishes at different strengths. This observation was utilized in the invention of a cell sorting system that sorts cells based on this adhesion strength difference. The resulting system is marketable at under $2000, compared to$50000 plus for current commercially available systems, and is the first commercially-oriented cell adhesion strength based cell sorter. The system allows for use of tissue culture flasks, a tool that any researcher using cells will be very familiar with, that have been customized with a removable bottom. After cells are adhered, the bottom is removed and placed into the cell sorting system and sealed with a removable sealing putty. Flow is imparted to the cells, generating a shear force over the surface of the cells, lifting the cells and depositing them into a centrifuge tube. Prior experiments have shown this system to have a cell purity and cell viability greater than currently available solutions without the use of expensive antibodies and using equipment available at a fraction of the cost.https://scholarscompass.vcu.edu/capstone/1145/thumbnail.jp

THE EFFECTS OF TERRESTRIAL AND EXTRATERRESTRIAL MECHANOSIGNALING ON CELLULAR AND TISSUE PROCESSES

Mechanobiological cues influence biological structures across all scales, from a single cell to a full body. These cues are vital for normal biological processes, such as cell migration and division, tissue structural composition, or astronaut orthostatic intolerance upon return to normal gravity. These cues also influence disease pathogenesis and progression, such as in cancer. Here, I will investigate mechanical influences on biological processes first at a single cell level, where we investigate force dynamics as a glioblastoma cell migrates and invades its surroundings. This investigation has impacts on high grade glioma invasion through the secondary structures of Scherer in the brain. This is a cancer type that has stubbornly resisted many treatment efforts and remains among the most lethal cancers with a 5-year survival of less than 10%. Next, we change scales to the tissue scale, where we investigate the effects of spaceflight on human engineered heart tissues. Spaceflight is well known to result in cardiovascular remodeling. This can have lethal consequences: the only astronauts to have left low Earth orbit are 5 times as likely to die of cardiovascular disease than the rest of the astronaut population. I show that spaceflight has negative consequences on engineered cardiac tissues, which may result from oxidative stress and mitochondrial dysfunction. Finally, both the spaceflight and cancer studies utilize polymeric substrates which are known to absorb drug compounds, preventing accurate drug screenings. We address this problem by showing that a polydimethylsiloxane-polyethylene glycol block copolymer can prevent drug absorption in engineered tissue systems, enabling future studies to conduct accurate drug screenings on the mechanobiological phenomena studied here. Together, this work has impacts on cancer migration, spaceflight, and microphysiological systems in general

Dynamic organelle distribution initiates actin-based spindle migration in mouse oocytes

© 2020, The Author(s). Migration of meiosis-I (MI) spindle from the cell center to a sub-cortical location is a critical step for mouse oocytes to undergo asymmetric meiotic cell division. In this study, we investigate the mechanism by which formin-2 (FMN2) orchestrates the initial movement of MI spindle. By defining protein domains responsible for targeting FMN2, we show that spindle-periphery localized FMN2 is required for spindle migration. The spindle-peripheral FMN2 nucleates short actin bundles from vesicles derived likely from the endoplasmic reticulum (ER) and concentrated in a layer outside the spindle. This layer is in turn surrounded by mitochondria. A model based on polymerizing actin filaments pushing against mitochondria, thus generating a counter force on the spindle, demonstrated an inherent ability of this system to break symmetry and evolve directional spindle motion. The model is further supported through experiments involving spatially biasing actin nucleation via optogenetics and disruption of mitochondrial distribution and dynamics

High-throughput longitudinal electrophysiology screening of mature chamber-specific hiPSC-CMs using optical mapping

Summary: hiPSC-CMs are being considered by the Food and Drug Administration and other regulatory agencies for in vitro cardiotoxicity screening to provide human-relevant safety data. Widespread adoption of hiPSC-CMs in regulatory and academic science is limited by the immature, fetal-like phenotype of the cells. Here, to advance the maturation state of hiPSC-CMs, we developed and validated a human perinatal stem cell-derived extracellular matrix coating applied to high-throughput cell culture plates. We also present and validate a cardiac optical mapping device designed for high-throughput functional assessment of mature hiPSC-CM action potentials using voltage-sensitive dye and calcium transients using calcium-sensitive dyes or genetically encoded calcium indicators (GECI, GCaMP6). We utilize the optical mapping device to provide new biological insight into mature chamber-specific hiPSC-CMs, responsiveness to cardioactive drugs, the effect of GCaMP6 genetic variants on electrophysiological function, and the effect of daily β-receptor stimulation on hiPSC-CM monolayer function and SERCA2a expression

CURRENT AND FUTURE GENETIC TECHNOLOGIES FOR FISHERIES AND AQUACULTURE: IMPLICATIONS FOR THE WORK OF FAO

Within the last few decades, advances in genetic technologies have created powerful and efficient tools for fisheries stock identification, genetic improvement and domestication of aquaculture species and characterization of changes in aquatic life due to environmental or anthropogenic influences. Emerging genetic tools are improving our understanding of organisms in aquatic ecosystems, in terms of diversity, distribution, abundance, movement, function and adaptation, and are being applied in aquaculture facilities and across fishery and aquaculture value chains. In this study expert elicitation was used to examine and predict current and potential future (10 year) impacts of the application of these novel technologies in fisheries and aquaculture. Highlighting the need to prepare sectors for likely changes that will follow. All suggestions received were collated into themes, to provide a conceptual framework that partitions potential impacts, and calls for required action — action required on governance, management and practical application of these innovations at both a national and international level.CURRENT AND FUTURE GENETIC TECHNOLOGIES FOR FISHERIES AND AQUACULTURE: IMPLICATIONS FOR THE WORK OF FAOpublishedVersio