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Biology subjects

Merry, C. L. R.

Publications and source records attributed to Merry, C. L. R..

3 recordsLinked to original sources

Species-specific WNT requirements shape somite morphogenesis in mammalian embryo models

Somitogenesis establishes the metameric body plan of the vertebrate embryo and can be recapitulated in vitro using stem cell-based embryo models. Although these systems are well established in individual species, they have rarely been compared across species, leaving unresolved how conserved developmental programmes generate species-specific tissue organisation. Here, we compare somitogenesis in embryo models derived from human, pig, sheep and cow pluripotent stem cells and investigate how WNT signalling and the extracellular matrix regulate axis segmentation. Under identical induction conditions, human and pig models diverge in cell-type composition and morphology, revealing distinct modes of axial elongation. Increasing WNT stimulation restores somite formation in pig models but fails to restore ordered segmentation. Furthermore, each of the four mammalian species requires a different level of WNT stimulation and matrix embedding time for somite formation. Finally, comparative analyses identify species-specific differences in progenitor dynamics. Human models, showing ordered segmentation, retain a posterior mesoderm-producing domain, whereas pig models show rapid depletion of this domain, associated with simultaneous somite formation. Together, these findings reveal species-specific requirements for mammalian somitogenesis in stem cell-derived embryo models and suggest that maintenance of a posterior axial domain contributes to ordered segmentation.

developmental biology↗

The development of ToF-SIMS for in-situ glycosaminoglycan analysis

Glycosaminoglycans (GAGs) are linear polysaccharides with essential roles in a myriad of biological processes. Despite their biological importance, methods to determine both spatial and compositional information is limited. Time-of-flight secondary ion mass spectrometry (ToF-SIMS) provides spatially resolved compositional information of biological molecules without enzymatic digestion or label incorporation, enabling unbiased analysis independent of enzyme or label selectivity, overcoming many current limitations in GAG analysis. Here, we present the identification and validation of GAG discriminatory ions from biological samples by comparison of spectra from purified GAGs and cells with genetically modified GAG biosynthetic pathways. Ions discriminatory of specific GAG sub-families are identified and related to GAG structural components. The analysis is applied to human induced pluripotent stem cells engineered to lack heparan sulphate (HS), where compensatory changes in GAG display that link to function were observed. Furthermore, the broad applicability and spatial resolution of the technique is highlighted through detection of a disease-induced reduction in HS within the individual glomeruli of diabetic mice.

biochemistry↗

OptoRheo: Simultaneous in situ micro-mechanical sensing and 3D imaging of live cell cultures

Biomechanical cues from the extracellular matrix (ECM) are essential for directing many cellular processes, from normal development and repair, to disease progression. To better understand cell-matrix interactions, we have developed a new instrument named OptoRheo that combines light sheet fluorescence microscopy with particle tracking microrheology. OptoRheo lets us image cells in 3D as they proliferate over several days while simultaneously sensing the mechanical properties of the surrounding extracellular and pericellular matrix at a sub-cellular length scale. OptoRheo can be used in two operational modalities (with and without an optical trap) to extend the dynamic range of microrheology measurements. We corroborated this by characterising the ECM surrounding live breast cancer cells in two distinct culture systems, cell clusters in 3D hydrogels and spheroids in suspension culture. This cutting-edge instrument will transform the exploration of drug transport through complex cell culture matrices and optimise the design of the next-generation of disease models.

bioengineering↗