A novel strategy to generate human iPSCs with enhanced mesodermal and endodermal differentiation capacity.

Induced pluripotent stem cells (iPSCs) have unlimited proliferative capacity and multi-lineage differentiation potential. Autologous iPSC-derived cells reduce the risk of immune rejection, making them promising resources for regenerative medicine. However, iPSCs derived using conventional methods often show heterogeneity and defects in differentiation potential in a subset of cells. To overcome these limitations, we developed a novel reprogramming strategy, termed the "4CL-primed" strategy, that incorporates a transient early-pluripotency (4CL) treatment during reprogramming. The 4CL-primed iPSCs retained the core characteristics of conventional iPSCs, including normal morphology, pluripotency marker expression, and karyotypic stability, while exhibiting enhanced mesodermal and endodermal differentiation capacity. Furthermore, the cells showed superior performance in directed pancreatic lineage differentiation, generating a higher proportion of pancreatic endocrine β-like cells. Collectively, the 4CL-primed strategy offers a more practical approach for mesodermal and endodermal differentiation research, particularly pancreatic lineage differentiation, thereby providing a more efficient source of pancreatic lineage cells for regenerative medicine and cell-based therapy.