The founder cells of the Nucleus pulposus, the centre of the intervertebral disc, originate in the embryonic notochord. After birth, mature notochordal cells (NC) are identified as key regulators of disc homeostasis. Better understanding of their biology has great potential in delaying the onset of disc degeneration or as a regenerative-cell source for disc repair. Using human pluripotent stem cells, we developed a two-step method to generate a stable NC-like population with a distinct molecular signature. Time-course analysis of lineage-specific markers shows that WNT pathway activation and transfection of the notochord-related transcription factor NOTO are sufficient to induce high levels of mesendoderm progenitors and favour their commitment toward the notochordal lineage instead of paraxial and lateral mesodermal or endodermal lineages. This study results in the identification of NOTO-regulated genes including some that are found expressed in human healthy disc tissue and highlights NOTO function in coordinating the gene network to human notochord differentiation.

Modelling rare neurogenetic diseases to develop new therapeutic strategies is highly challenging. The use of human-induced pluripotent stem cells (hiPSCs) is a powerful approach to obtain specialized cells from patients. For hereditary peripheral neuropathies, such as Charcot–Marie–Tooth disease (CMT) Type II, spinal motor neurons (MNs) are impaired but are very difficult to study. Although several protocols are available to differentiate hiPSCs into neurons, their efficiency is still poor for CMT patients. Thus, our goal was to develop a robust, easy, and reproducible protocol to obtain MNs from CMT patient hiPSCs. The presented protocol generates MNs within 20 days, with a success rate of 80%, using specifically chosen molecules, such as Sonic Hedgehog or retinoic acid. The timing and concentrations of the factors used to induce differentiation are crucial and are given hereby. We then assessed the MNs by optic microscopy, immunocytochemistry (Islet1/2, HB9, Tuj1, and PGP9.5), and electrophysiological recordings. This method of generating MNs from CMT patients in vitro shows promise for the further development of assays to understand the pathological mechanisms of CMT and for drug screening.

Background: The constant emergence of novel psychoactive substances (NPS) poses significant analytical challenges due to their structural diversity, low concentrations in complex biological matrices, and the limited availability of certified reference materials. This review summarizes current and emerging analytical strategies for detecting novel substances and evaluates the contribution of innovative technologies for toxicological surveillance. Methods: Studies addressing analytical techniques for NPS detection including chromatographic, spectrometric, ambient ionization, and artificial intelligence (AI)-assisted methods were systematically reviewed, with emphasis on applicability, sensitivity, and adaptability in clinical, forensic, and environmental contexts. Results: Conventional techniques such as liquid chromatography coupled with tandem mass spectrometry and immunoassays provide reliable detection of known compounds but are limited by their targeted scope for detecting newly synthesized analogs. High-resolution mass spectrometry, nuclear magnetic resonance, and direct ionization approaches offer broader detection capabilities but remain resource-intensive. Recent advances in AI—including spectral prediction, structural classification, and de novo compound generation—offer promising new avenues for untargeted screening and early compound identification. In addition, emerging tools such as molecular networking, wastewater-based epidemiology, and portable mass spectrometry devices further enhance real-time monitoring and surveillance capacity. Conclusions: An integrated, adaptive analytical framework combining high-resolution mass spectrometry, ambient ionization, and AI-driven data analysis will help address the fast-evolving NPS landscape. Standardization of reference materials, harmonization across laboratories, and interdisciplinary collaboration between chemists, clinicians, and regulatory agencies will be key to improving the speed, sensitivity, and public health impact of NPS detection.

Membrane transporters play a central role in the absorption, distribution, metabolism, and elimination of drugs. Beyond drug handling, these transporters regulate the transport of endogenous metabolites, contributing to physiological homeostasis. Their activity is regulated not only by intrinsic cellular mechanisms but also by signals from distant organs. Understanding these processes is crucial for accurately predicting both local and systemic pharmacokinetics, as transporter function influences drug concentrations and effects at target tissues and throughout the organism. However, most current in vitro models do not recapitulate interorgan interactions or the dynamic regulation of transporter function. Microphysiological systems, particularly organ-on-chip, offer a promising approach to studying transporter biology in a more integrated and physiologically relevant context. This review provides an overview of the development and application of organ-on-chip technologies to investigate drug transporters and their regulation. Although significant technical progress has been made, these systems often remain focused on engineering or industrial goals, with limited impact on fundamental biological questions. Moreover, the high degree of customization and widespread use of homemade organ-on-chip models, with diverse materials, geometries, and cell sources, have created substantial barriers to reproducibility and cross-laboratory comparison. We here assess the current state of the field and identify gaps in modeling transporter regulation and interorgan communication. We also explore the potential of organ-on-chip to advance our understanding of systemic pharmacology. This review aims to reposition organ-on-chip platforms not only as technological demonstrators but also as experimental systems capable of generating new biological insights into transporter function and organ crosstalk. SIGNIFICANCE STATEMENT: Membrane transporters are central regulators of drug disposition and systemic homeostasis, yet their interorgan regulation remains poorly captured by conventional models. This review highlights how organ-on-chip technologies can bridge this gap, enabling mechanistic investigation of transporter function and advancing predictive, human-relevant pharmacology.

Peripheral neuropathies are disorders affecting the peripheral nervous system. Among them, Charcot-Marie-Tooth disease is an inherited sensorimotor neuropathy for which no effective treatment exists yet. Research on Charcot-Marie-Tooth disease has been hampered by difficulties in accessing relevant cells, such as sensory and motor neurons, Schwann cells, and myocytes, which interact at the neuromuscular junction, the specialized synapses formed between nerves and skeletal muscles. This review first outlines the various in vivo models and methods used to study neuromuscular junction deficiencies in Charcot-Marie-Tooth disease. We then explore novel in vitro techniques and models, including complex hiPSC-derived cultures, which offer promising isogenic and reproducible neuromuscular junction models. The adaptability of in vitro culture methods, including cell origin, cell-type combinations, and choice of culture format, adds complexity and excitement to this rapidly evolving field. This review aims to recapitulate available tools for studying Charcot-Marie-Tooth disease to understand its pathophysiological mechanisms and test potential therapies.