A5012356388- 3D Printing in Biomedical Research
- Pluripotent Stem Cells Research
- Innovative Microfluidic and Catalytic Techniques Innovation
- Pancreatic function and diabetes
- Single-cell and spatial transcriptomics
- Cell Image Analysis Techniques
- CRISPR and Genetic Engineering
- Renal and related cancers
- Tissue Engineering and Regenerative Medicine
- Liver physiology and pathology
- RNA Research and Splicing
- Monoclonal and Polyclonal Antibodies Research
- Microfluidic and Bio-sensing Technologies
- SARS-CoV-2 and COVID-19 Research
- RNA modifications and cancer
- Transgenic Plants and Applications
- Genetic and Kidney Cyst Diseases
- Electrowetting and Microfluidic Technologies
- Pregnancy and preeclampsia studies
- Prenatal Screening and Diagnostics
- Mesenchymal stem cell research
- Cancer-related molecular mechanisms research
University of Cambridge
2020-2024
Wellcome/MRC Cambridge Stem Cell Institute
2020-2023
Medical Research Council
2020-2023
Abstract Most methods for single-cell transcriptome sequencing amplify the termini of polyadenylated transcripts, capturing only a small fraction total cellular transcriptome. This precludes detection many long non-coding, short non-coding and non-polyadenylated protein-coding transcripts hinders alternative splicing analysis. We, therefore, developed VASA-seq to detect in single cells, which is enabled by fragmenting tailing all RNA molecules subsequent cell lysis. The method compatible...
Abstract Droplet microfluidic methods have massively increased the throughput of single-cell sequencing campaigns. The benefit scale-up is, however, accompanied by background noise when processing challenging samples and overall RNA capture efficiency is lower. These drawbacks stem from lack strategies to enrich for high-quality material or specific cell types at moment encapsulation absence implementable multi-step enzymatic processes that increase capture. Here we alleviate both...
In the liver, ductal cells rarely proliferate during homeostasis but do so transiently after tissue injury. These can be expanded as organoids that recapitulate several of cell-autonomous mechanisms regeneration lack stromal interactions native tissue. Here, using organoid co-cultures ductal-to-mesenchymal cell architecture portal tract, we demonstrate a subpopulation mouse periportal mesenchymal exerts dual control on proliferation epithelium. Ductal is either induced and sustained or,...
Abstract Monoclonal antibodies are increasingly used to prevent and treat viral infections pivotal in pandemic response efforts. Antibody-secreting cells (ASCs; plasma plasmablasts) an excellent source of high-affinity with therapeutic potential. Current methods study antigen-specific ASCs either have low throughput, require expensive labor-intensive screening or technically demanding therefore not widely accessible. Here we present a straightforward technology for the rapid discovery...
Human periimplantation development requires the transformation of naive pluripotent epiblast into a polarized epithelium. Lumenogenesis plays critical role in this process, as undergoes rosette formation and lumen expansion to form amniotic cavity. Here, we present high-throughput vitro model for morphogenesis. We established microfluidic workflow encapsulate human stem cells (hPSCs) monodisperse agarose microgels. Strikingly, hPSCs self-organized spheroids that could be maintained...
Combining live imaging with the ability to retrieve individual cells of interest remains a technical challenge. precise cell retrieval is particular when studying highly dynamic or transient, asynchronous, heterogeneous biological and developmental processes. Here, we present method encapsulate in 3D hydrogel matrix, via bead compartmentalisation. Using small-scale screen, optimised matrix conditions for culture multilineage differentiation mouse embryonic stem cells. Moreover, designed...
The early specification and rapid growth of extraembryonic membranes are distinctive hallmarks primate embryogenesis. These complex tasks resolved through an intricate combination signals controlling the induction lineages and, at same time, safeguarding pluripotent epiblast. Here, we delineate orchestrating epiblast amnion identity. We encapsulated marmoset stem cells into agarose microgels identified culture conditions for development epiblast- amnion-spheroids. Spatial identity mapping...
The assembly of robust, modular biological components into complex functional systems is central to synthetic biology. Here, we apply "plug and play" design principles a solid-phase protein display system that facilitates purification assays. Specifically, capture proteins on polyacrylamide hydrogel beads (PHD beads) made in microfluidic droplet generators. These monodisperse PHD are decorated with predefined amounts anchors, methacrylate-PEG-benzylguanine (BG) methacrylate-PEG-chloroalkane...
Abstract Biomechanical cues are instrumental in guiding embryonic development and cell differentiation. Understanding how these physical stimuli translate into transcriptional programs will provide insight mechanisms underlying mammalian pre-implantation development. Here, we explore this type of regulation by exerting microenvironmental control over mouse stem cells. Microfluidic encapsulation cells agarose microgels stabilizes the naive pluripotency network specifically induces expression...
ABSTRACT In recent years, single-cell transcriptome sequencing has revolutionized biology, allowing for the unbiased characterization of cellular subpopulations. However, most methods amplify termini polyadenylated transcripts capturing only a small fraction total transcriptome. This precludes detection many long non-coding, short non-coding and non-polyadenylated protein-coding transcripts. Additionally, workflows do not sequence full transcript hindering analysis alternative splicing. We...
Abstract Monoclonal antibodies are increasingly used to prevent and treat viral infections, playing a pivotal role in pandemic response efforts. Antibody secreting cells (ASCs, plasma plasmablasts) an excellent source of high-affinity with therapeutic potential. Current methodologies study antigen-specific ASCs either have low throughput, require expensive labour-intensive screening or technically demanding therefore not accessible the wider research community. Here, we present...
Abstract Droplet microfluidic methods have massively increased the throughput of single-cell sequencing campaigns. The benefit scale-up is, however, accompanied by background noise when processing challenging samples and overall RNA capture efficiency is lower. These drawbacks stem from lack strategies to enrich for high-quality material or specific cell types at moment encapsulation absence implementable multi-step enzymatic processes that increase capture. Here we alleviate both...
Within the peri-portal region of adult liver, portal fibroblasts exist in close proximity to epithelial ductal/cholangiocyte cells. However, cellular interactions between them are poorly understood. Here, we provide two co-culture techniques incorporate liver mesenchyme into ductal cell organoids, which recapitulate aspects their vitro. We integrate several from isolation and expansion by microfluidic co-encapsulation or 2D-Matrigel layer. The protocol is easily adaptable other cells organs....
ABSTRACT Combining live imaging with the ability to retrieve individual cells of interest remains a technical challenge. These combined methods are particular when studying highly dynamic or transient, asynchronous heterogeneous cell biological and developmental processes. Here we present method encapsulate in 3D hydrogel matrix, via droplet compartmentalisation. Using small-scale screen, optimised matrix conditions for culture multilineage differentiation mouse embryonic stem (ES) cells....
Abstract Biomechanical cues are instrumental in guiding embryonic development and cell differentiation. Understanding how these physical stimuli translate into transcriptional programs could provide insight mechanisms underlying mammalian pre-implantation development. Here, we explore this by exerting microenvironmental control over mouse stem cells (ESCs). Microfluidic encapsulation of ESCs agarose microgels stabilized the naïve pluripotency network specifically induced expression...
Abstract In the homeostatic liver, ductal cells intermingle with a microenvironment of endothelial and mesenchymal to form functional unit portal tract. Ductal proliferate rarely in homeostasis but do so transiently after tissue injury replenish any lost epithelium. We have shown that liver can be expanded as organoids recapitulate several cell-autonomous mechanisms regeneration, lack stromal cell milieu biliary tract vivo . Here, we describe subpopulation SCA1 + periportal closely surrounds...