The sympathetic nervous system densely innervates all cardiac chambers and is a key player in cardiac control, yet this relationship has scarcely been investigated using a stem cell-based model. This study investigates the effects that sympathetic neurons (SNs) have on human pluripotent stem cell-derived cardiomyocytes (hPSC-CMs) in vitro, and whether they induce any degree of functional or structural maturity in these conventionally immature cells. SNs were isolated from neonatal rat pups, and cocultured with hPSC-CMs for up to 15 days. Structural changes in hPSC-CMs were analysed by microscopy techniques. Fluorescence resonance energy transfer was used to measure second messenger molecule cAMP production and β-adrenergic receptor (βAR) response. Contractile and Ca2+ transient activity was measured using CytoCypher. These cocultures promoted hPSC-CM structural elongation and increased sarcomere organization. Furthermore, the βAR response of cocultured hiPSC-CMs was larger, indicated by increased cAMP production upon neuronal nicotinic stimulation. Faster contraction and ratiometric Ca2+ transient peak height and kinetic parameters strongly indicate increased chronotropic response in coculture. Coculture with SNs also elicited an increase in action potential amplitude and depolarization velocity, further confirming that SNs contribute to hiPSC-CM functional maturation. Overall, we have found that SNs modulate hPSC-CMs in vitro, inducing a more mature functional response. As an in vitro tool, these cocultures could serve as a model of sympathoadrenergic disease, enabling new discovery avenues. KEY POINTS: The sympathetic nervous system controls the involuntary 'fight-or-flight' response, with the heart being one of key target organs. In certain neuro-cardiac diseases, the input from the sympathetic nervous system is hyperregulated, and can lead to increased speed or force of the heart's contraction. Human induced pluripotent stem cells (hiPSCs) represent a rapidly evolving field which allow us to create a cell of interest and model its structural and functional activity in a dish. Here we have created hiPSC-derived cardiomyocytes (hiPSC-CMs) and cocultured them with sympathetic neurons (SNs). We found that SNs are able to modulate structure of the hiPSC-CMs by reducing their circularity and increasing sarcomeric organization, and can significantly increase the speed of contraction and Ca2+ handling. Together, our data provide a platform to investigate the neuro-cardiac relationship in vitro, which could be used for patient-specific disease modelling in future.
© 2025 The Author(s). The Journal of Physiology published by John Wiley & Sons Ltd on behalf of The Physiological Society.

Readily accessible human pancreatic beta cells that are functionally close to primary adult beta cells are a crucial model to better understand human beta cell physiology and develop new treatments for diabetes. We here report the characterization of EndoC-βH5 cells, the latest in the EndoC-βH cell family.
EndoC-βH5 cells were generated by integrative gene transfer of immortalizing transgenes hTERT and SV40 large T along with Herpes Simplex Virus-1 thymidine kinase into human fetal pancreas. Immortalizing transgenes were removed after amplification using CRE activation and remaining non-excized cells eliminated using ganciclovir. Resulting cells were distributed as ready to use EndoC-βH5 cells. We performed transcriptome, immunological and extensive functional assays.
Ready to use EndoC-βH5 cells display highly efficient glucose dependent insulin secretion. A robust 10-fold insulin secretion index was observed and reproduced in four independent laboratories across Europe. EndoC-βH5 cells secrete insulin in a dynamic manner in response to glucose and secretion is further potentiated by GIP and GLP-1 analogs. RNA-seq confirmed abundant expression of beta cell transcription factors and functional markers, including incretin receptors. Cytokines induce a gene expression signature of inflammatory pathways and antigen processing and presentation. Finally, modified HLA-A2 expressing EndoC-βH5 cells elicit specific A2-alloreactive CD8 T cell activation.
EndoC-βH5 cells represent a unique storable and ready to use human pancreatic beta cell model with highly robust and reproducible features. Such cells are thus relevant for the study of beta cell function, screening and validation of new drugs, and development of disease models.
Copyright © 2023 The Authors. Published by Elsevier GmbH.. All rights reserved.

Decreased left ventricle (LV) function caused by genetic mutations or injury often leads to debilitating and fatal cardiovascular disease. LV cardiomyocytes are, therefore, a potentially valuable therapeutical target. Human pluripotent stem cell-derived cardiomyocytes (hPSC-CMs) are neither homogeneous nor functionally mature, which reduces their utility. Here, we exploit cardiac development knowledge to instruct differentiation of hPSCs specifically toward LV cardiomyocytes. Correct mesoderm patterning and retinoic acid pathway blocking are essential to generate near-homogenous LV-specific hPSC-CMs (hPSC-LV-CMs). These cells transit via first heart field progenitors and display typical ventricular action potentials. Importantly, hPSC-LV-CMs exhibit increased metabolism, reduced proliferation, and improved cytoarchitecture and functional maturity compared with age-matched cardiomyocytes generated using the standard WNT-ON/WNT-OFF protocol. Similarly, engineered heart tissues made from hPSC-LV-CMs are better organized, produce higher force, and beat more slowly but can be paced to physiological levels. Together, we show that functionally matured hPSC-LV-CMs can be obtained rapidly without exposure to current maturation regimes.
Crown Copyright © 2023.

Circulating tumor cells (CTCs) have been identified as responsible for the spread of tumors to other organs of the body. In this sense, the development of sensitive and specific assays for their detection is important to reduce the number of deaths due to metastases. Here, we assessed whether the detection of CTCs in peripheral blood can serve in the construction of a panel of diagnosis and monitoring treatments of breast cancer (BC), focusing on the expression of markers of epithelial-mesenchymal transition. Through analyzing the blood from women without breast alterations (control), women with benign alterations, women with breast cancer without chemotherapy, and women with breast cancer with chemotherapy, we identified the best markers by transcriptional levels and determined three profiles of CTCs (mesenchymal, intermediate, and epithelial) by flow cytometry which, combined, can be used for diagnosis and therapy monitoring with sensitivity and specificity between 80% and 100%. Therefore, we have developed a method for detecting breast cancer based on the analysis of CTC profiles by epithelial-mesenchymal transition markers which, combined, can be used for the diagnosis and monitoring of therapy.

The HNF1αp291fsinsC truncation is the most common mutation associated with maturity-onset diabetes of the young 3 (MODY3). Although shown to impair HNF1α signaling, the mechanism by which HNF1αp291fsinsC causes MODY3 is not fully understood. Here we use MODY3 patient and CRISPR/Cas9-engineered human induced pluripotent stem cells (hiPSCs) grown as 3D organoids to investigate how HNF1αp291fsinsC affects hiPSC differentiation during pancreatic development. HNF1αp291fsinsC hiPSCs shows reduced pancreatic progenitor and β cell differentiation. Mechanistically, HNF1αp291fsinsC interacts with HNF1β and inhibits its function, and disrupting this interaction partially rescues HNF1β-dependent transcription. HNF1β overexpression in the HNF1αp291fsinsC patient organoid line increases PDX1+ progenitors, while HNF1β overexpression in the HNF1αp291fsinsC patient iPSC line partially rescues β cell differentiation. Our study highlights the capability of pancreas progenitor-derived organoids to model disease in vitro. Additionally, it uncovers an HNF1β-mediated mechanism linked to HNF1α truncation that affects progenitor differentiation and could explain the clinical heterogeneity observed in MODY3 patients.Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.