Biology subjects

Priest, C.

Publications and source records attributed to Priest, C..

2 recordsLinked to original sources

bioRxiv2024

Human stem cell-derived GABAergic interneuron development reveals early emergence of subtype diversity followed by gradual electrochemical maturation

Medial ganglionic eminence-derived inhibitory GABAergic pallial interneurons (MGE-pINs) are essential regulators of cortical circuits; their dysfunction is associated with numerous neurological disorders. We developed human (h) MGE-pINs from pluripotent stem cells for the treatment of drug-resistant epilepsy. Here, we analyzed xenografted hMGE-pINs over the lifespan of host mice using single nuclei RNA sequencing. Comparative transcriptomics against endogenous human brain datasets revealed that 97% of grafted cells developed into somatostatin (SST) and parvalbumin (PVALB) subtypes, including populations that exhibit selective vulnerability in Alzheimers disease. Transplanted hMGE-pINs demonstrated rapid emergence of subclass features, progressing through distinct transcriptional states sequentially involving neuronal migration, synapse organization, and membrane maturation. We present molecular, electrophysiological, and morphological data that collectively confirm the derivation of diverse bona-fide human SST and PVALB subtypes, providing a high-fidelity model to study human MGE-pIN development and functional maturation as well as a compositional atlas for regenerative cell therapy applications.

neuroscience↗

bioRxiv2018

Intracellular delivery of mRNA to human primary T cells with microfluidic vortex shedding

Intracellular delivery of functional macromolecules, such as DNA and RNA, across the cell membrane and into the cytosol, is a critical process in both biology and medicine. Herein, we develop and use microfluidic chips containing post arrays to induce microfluidic vortex shedding, or VS, for cell membrane poration that permits delivery of mRNA into primary human T lymphocytes. We demonstrate transfection with VS by delivery of a 996-nucleotide mRNA construct encoding enhanced green fluorescent protein (EGFP) and assessed transfection efficiencies by quantifying levels of EGFP protein expression. We achieved high transfection efficiency (63.6 {+/-} 3.44% EGFP+ viable cells) with high cell viability (77.3 {+/-} 0.58%) and recovery (88.7 {+/-} 3.21%) in CD3+ T cells 19 hrs after VS processing. Importantly, we show that processing cells via VS does not negatively affect cell growth rates or alter cell states. We also demonstrate processing speeds of greater than 2.0 x 106 cells s-1 at volumes ranging from 0.1 to 1.5 milliliters. Altogether, these results highlight the use of VS as a rapid and gentle delivery method with promising potential to engineer primary human cells for research and clinical applications.

bioengineering↗