Biology subjects

Berry, C. C.

Publications and source records attributed to Berry, C. C..

4 recordsLinked to original sources

bioRxiv2026

Engineering tunable synthetic fibrotic niches to model acinar-to-ductal metaplasia in 2D pancreatic culture systems

Acinar-to-ductal metaplasia (ADM) is an early pancreatic cell-state transition associated with injury, inflammation, and progression toward pancreatic intraepithelial neoplasia (PanIN). Although genetic and inflammatory drivers of ADM are well established, the contribution of extracellular-matrix mechanics is difficult to isolate because tissue stiffening is accompanied by changes in matrix composition and soluble signaling. Here, we describe a two-dimensional polyacrylamide (PAA) hydrogel platform that independently varies substrate stiffness and adhesive ligand presentation for murine pancreatic acinar 266-6 cells. Hydrogels approximate soft pancreatic tissue (0.50 kPa), a transitional mechanical condition (4.48 kPa), and fibrotic tissue (18.29 kPa). RGD provides a conventional adhesive cue relevant across healthy-to-fibrotic extracellular-matrix states, whereas incorporation of laminin-derived isoleucine-lysine-valine-alanine-valine (IKVAV) creates a more basement-membrane-like environment consistent with preservation of acinar identity. The platform was also adapted to include gelatin, a collagen-derived protein used here to model a more collagen-rich fibrotic matrix context. Peptide coupling is calibrated using fluorescently labeled RGD, and responses after 3 days are assessed by morphology, alpha-amylase 2 (AMY2), Yes-associated protein (YAP) nuclear-to-cytoplasmic localization, and sex-determining region Y-box transcription factor 9 (SOX9). Increasing stiffness increased nuclear YAP localization and reduced AMY2 expression. IKVAV-containing conditions supported greater attachment on softer matrices and partially preserved acinar-like phenotype on stiff gels despite elevated nuclear YAP. Across all ligand conditions, increasing stiffness reduced roundness and geometric regularity/solidity and increased aspect ratio; on gelatin-modified hydrogels, these phenotypic changes corresponded to increased cell-area-normalized SOX9 fluorescence. Together, these readouts identify a stiffness-dependent shift from an acinar-associated state toward an ADM-associated ductal phenotype, while ligand context modulates how this mechanical response is expressed. This protocol provides a controllable, adaptable system for separating mechanical and biochemical extracellular-matrix inputs during pancreatic phenotypic plasticity.

cancer biology↗

bioRxiv2022

Extracellular vesicle cargo metabolome changes in response to the mesenchymal stromal cell microenvironment and influences cell quiescence and activation in a human breast cancer cell model

Breast cancer is the leading cause of cancer mortality in women worldwide and commonly metastasizes to the bone marrow, drastically reducing patient prognosis and survival. In the bone marrow niche, metastatic cells can enter into a dormant state, thereby evading immune surveillance and treatment, and can be reactivated to enter a proliferative state due to poorly understood cues. Mesenchymal stromal cells (MSCs) maintain cells in this niche partly by secreting extracellular matrix and paracrine factors and by responding to regenerative cues. MSCs also produce extracellular vesicles (EVs) that carry a range of cargoes, some of which are implicated in cell signalling. Here, we investigate if the changing metabolic state of MSCs alters the cargoes they package into EVs, and how these changing cargoes act on dormant breast cancer cells (BCCs) using an in vitro BCC spheroid model and a scratch assay to create a regenerative demand on MSCs. Our findings show that EVs produced by standard MSCs contain glycolytic metabolites that maintain BCC dormancy. When MSCs are placed under a regenerative demand and increase their respiration to fuel differentiation, these metabolites disappear from the EV cargo and their absence encourages rapid growth in the BCC spheroids. This work implicates EVs in cancer cell dormancy in the bone marrow niche and indicates that pressures on the niche, such as regeneration, can be a driver of BCC activation.

cell biology↗

bioRxiv2017

Temporal differentiation of bovine airway epithelial cells grown at an air-liquid interface

The respiratory epithelium is exposed to assault by toxins and pathogens through the process of inhalation, which has numerous implications on both human and animal health. As such, there is a need to develop and characterise an in vitro model of the airway epithelium to study respiratory pathologies during infection or toxicology experiments. This has been achieved by growing airway epithelial cells at an air-liquid interface (ALI). Characterisation of ALI models are not well-defined for airway epithelial cells derived from non-human species. In this study we have fully characterised a bovine airway epithelial cell models (AECM) grown at an ALI in relation to ex vivo tissue. The morphology of the model was monitored at three day intervals, to identify the time-period at which the culture was optimally differentiated. The model was shown to be fully-differentiated by day 21 post-ALI. The culture formed a stereotypical pseudostratified, columnar epithelium containing the major cell types of the bronchial epithelium (ciliated-, goblet- and basal cells). Once fully differentiated the bovine AECM displayed both barrier function, through the formation of tight-junctions, and active mucociliary clearance, important properties of the mucosal barrier. The bovine bronchial epithelial cells remained stable for three weeks, with no evidence of deterioration or dedifferentiation. The window in which the model displayed full differentiation was determined to be between day 21-42 post-ALI. Through comparison with ex vivo tissue derived from donor animals, our bovine AECM was shown to be highly representative of the in vivo bovine bronchial epithelium and can be utilised in the study of respiratory pathologies.

microbiology↗

bioRxiv2017

Interactions of pathogenic and commensal strains of Mannheimia haemolytica with differentiated bovine airway epithelial cells grown at an air-liquid interface

Mannheimia haemolytica serotype A2 is a common commensal species present in the nasopharynx of healthy cattle. However, prior to the onset of bovine pneumonic pasteurellosis, there is sudden increase in M. haemolytica serotype A1 within the upper respiratory tract. The events during this selective proliferation of serotype A1 strains are poorly characterised. In this investigation, a differentiated bovine airway epithelial cell culture was used to study the interactions of A1 and A2 bovine isolates with the respiratory epithelium. This model reproduced the key defences of the airway epithelium, including tight junctions and mucociliary clearance. Although initial adherence of the serotype A1 strains was low, by 12 hours post-infection the bacteria was able to traverse the tight junctions to form foci of infection below the apical surface. The size, density and number of these foci increased with time, as did the cytopathic effects observed in the bovine bronchial epithelial cells. Penetration of M. haemolytica A1 into the sub-apical epithelium was shown to be through transcytosis but not paracytosis. Commensal A2 bovine isolates however were not capable of colonising the model to a high degree, and did not penetrate the epithelium following initial adherence at the apical surface. This difference in their ability to colonise the respiratory epithelium may account for the sudden proliferation of serotype A1 in the onset of pneumonia pasteurellosis. The pathogenesis observed was replicated by virulent A2 ovine isolates; however colonisation was 10-fold lower in comparison to bovine A1 strains. This investigation provides new insight into the interactions of M. haemolytica with bovine airway epithelial cells which are occurring in vivo during pneumonia pasteurellosis.

microbiology↗