From hPSCs to MSCs: differentiation strategies, pathways, and the emergence of common regulatory networks

Mesenchymal stem/stromal cells (MSCs) derived from human pluripotent stem cells (hPSCs) represent a scalable and homogeneous source for regenerative medicine. To date, multiple differentiation protocols have been developed to direct hPSCs toward an MSC fate, with intermediate cell states arising from diverse lineages, including trophoblast, neural crest, mesoderm, and endoderm. Despite these divergent differentiation strategies, the induced MSCs exhibit similar phenotypes and biological functions, suggesting convergent molecular programs underlying MSC specification. In this review, we discuss current strategies for differentiating hPSCs into MSCs and summarize the key signaling pathways, with a focus on the transcriptional regulators that govern these lineage-specific differentiation routes. To identify common regulatory nodes across different lineages, we analyzed publicly available transcriptomic datasets from representative hPSC-to-MSC protocols deposited in the Gene Expression Omnibus (GEO) database. Comparative analysis revealed a core set of consistently dysregulated genes and enriched pathways, particularly those involved in extracellular matrix (ECM)-receptor interaction, focal adhesion, and the PI3K–Akt signaling pathway. Notably, SMAD3, along with AP-1 family members (JUN, JUND, FOSL1, FOSL2) and the associated regulatory targets (FN1 and COL1A1) emerged as recurrent hubs in mesenchymal commitment. These findings highlight both the plasticity and convergence in the induction of MSCs from hPSCs and provide a molecular framework for optimizing differentiation strategies and ensuring product consistency in regenerative applications.