Vertti-Quintero Nadia, Delahousse Clara, Aristov Andrey, Traboulsi Tatiana, Cossec Jack-Christophe, Baroud Charles N, Sart Sébastien
Physical Microfluidics and Bioengineering Unit, Institut Pasteur, Université Paris Cité, 25-28 Rue du Dr. Roux, Paris, 75015, France.
Nuclear Organization and Oncogenesis Unit, Institut Pasteur, Université Paris Cité, 25-28 Rue du Dr. Roux, Paris, 75015, France.
Stem Cell Res Ther. 2025 Sep 26;16(1):510. doi: 10.1186/s13287-025-04625-7.
The differentiation of pluripotent stem cells (PSCs) and their self-organization into organoids are influenced by cell-cell interactions mediated by contacts and secreted molecules. These interactions are enhanced in microfluidic droplets due to confinement and small culture volumes. However, a comprehensive study on the culture of PSCs within droplets and the impact of this microenvironment has yet to be conducted.
In this study, we present a droplet platform for the 3D culture of PSCs at various stages of cellular commitment, such as pluripotency, spontaneous differentiation and directed differentiation towards gastruloids and cardioids.
We demonstrate PSC differentiation into the three germ layers and the feasibility of organoid formation within droplets. Our findings reveal that culturing PSCs in confined volumes regulates cell fate decisions, promoting tissue patterning in gastruloids through the sequential induction of growth and migration of distinct differentiated cell populations, and facilitating the self-organization of cardiac organoids.
This technological approach provides unique insights into the intrinsic factors regulating tissue self-patterning in vitro, with potential applications in drug screening and disease modeling.
多能干细胞（PSC）的分化及其自组织形成类器官受到由细胞接触和分泌分子介导的细胞间相互作用的影响。由于受限和小培养体积，这些相互作用在微流控液滴中得到增强。然而，尚未对液滴内PSC的培养及其微环境的影响进行全面研究。
在本研究中，我们提出了一个液滴平台，用于在细胞定向的各个阶段对PSC进行三维培养，如多能性、自发分化以及向原肠胚样和心脏样细胞的定向分化。
我们证明了PSC在液滴内分化为三个胚层以及形成类器官的可行性。我们的研究结果表明，在受限体积中培养PSC可调节细胞命运决定，通过依次诱导不同分化细胞群体的生长和迁移促进原肠胚样细胞中的组织模式形成，并促进心脏类器官的自组织。
这种技术方法为体外调节组织自模式形成的内在因素提供了独特见解，在药物筛选和疾病建模方面具有潜在应用。
