Summary: To investigate the co-development of vasculature, mesenchyme, and epithelium crucial for organogenesis and the acquisition of organ-specific characteristics, we constructed a human pluripotent stem cell-derived organoid system comprising lung or intestinal epithelium surrounded by organotypic mesenchyme and vasculature. We demonstrated the pivotal role of co-differentiating mesoderm and endoderm via precise BMP regulation in generating multilineage organoids and gut tube patterning. Single-cell RNA-seq analysis revealed organ specificity in endothelium and mesenchyme, and uncovered key ligands driving endothelial specification in the lung (e.g., WNT2B and Semaphorins) or intestine (e.g., GDF15). Upon transplantation under the kidney capsule in mice, these organoids further matured and developed perfusable human-specific sub-epithelial capillaries. Additionally, our model recapitulated the abnormal endothelial-epithelial crosstalk in patients with FOXF1 deletion or mutations. Multilineage organoids provide a unique platform to study developmental cues guiding endothelial and mesenchymal cell fate determination, and investigate intricate cell-cell communications in human organogenesis and disease. Highlights: BMP signaling fine-tunes the co-differentiation of mesoderm and endoderm. The cellular composition in multilineage organoids resembles that of human fetal organs. Mesenchyme and endothelium co-developed within the organoids adopt organ-specific characteristics. Multilineage organoids recapitulate abnormal endothelial-epithelial crosstalk in FOXF1-associated disorders.

Lung cancer remains the leading cause of cancer-related mortality worldwide, with non-small cell lung cancer accounting for a majority of cases. Despite advances in targeted therapies and immunotherapy, challenges such as tumor heterogeneity, resistance mechanisms, and limited preclinical models hinder treatment efficacy. Traditional cancer models, including 2D cell cultures and animal models, often fail to accurately replicate the lung's complex architecture, microenvironment, and biomechanical cues, leading to poor predictive performance in drug development. Microfluidic-based organ-on-a-chip technology offers a promising alternative by integrating human-derived cells with precisely controlled perfusion, mechanical cues, and tumor–stroma interactions in physiologically relevant 3D models. These platforms enable the study of lung cancer biology, drug responses, and patient-specific therapeutic outcomes with improved accuracy. In this review, we discuss recent advancements in microfluidic systems for recapitulating normal lung physiology and 3D lung cancer microenvironment, covering various microfluidic platforms with applications in disease modeling and drug testing. Unlike other review articles, we bring first-hand insights from clinicians about the current treatment practice for lung cancer and the clinical utilities of lung cancer-on-a-chip models, which bioengineers have been seeking. We also highlight the translational potential of these systems in personalized oncology and the need for interdisciplinary collaborations, particularly with clinicians, to enhance their clinical impact.

Endocardium lines the inner layer of the heart ventricle and serves as the source of valve endothelial cells and interstitial cells. Previously, endocardium-associated abnormalities in hypoplastic left heart syndrome (HLHS) have been reported, including endocardial fibroelastosis (EFE) and mitral and aortic valve malformation. However, few mechanistic studies have investigated the molecular pathological changes in endocardial cells. Recently, the emergence of a powerful in vitro system—induced pluripotent stem cells (iPSCs)—was applied to study various genetic diseases, including HLHS. This review summarized current in vitro studies in understanding the endocardial pathology in HLHS, emphasizing new findings of the cellular phenotypes and underlying molecular mechanisms. Lastly, a future perspective is provided regarding the better recapitulation of endocardial phenotypes in a dish.