Pluripotency-Preserving Culture System for Pancreatic Stellate Cells Using MSC Medium: From Quiescence Maintenance to Regenerative Potential.

Abstract

Introduction: Pancreatic stellate cells (PSCs) play a central role in pancreatic physiology and disease, and the transition between their quiescent and activated states influences processes such as fibrosis and regeneration. However, the stem cell-like properties of PSCs remain unclear. This study aims to clarify the stem cell characteristics of PSCs and establish a method to maintain their quiescent state, thereby providing a new perspective on pancreatic regeneration.

Methods: PSCs from Lrat-cre; Rosa26-tdTomato mice were fate-traced, and immunofluorescence was used to assess Lrat, Oct4, and Nanog co-localization. Primary PSCs from C57BL/6 mice were cultured in mesenchymal stem cell (MSC) or DMEM/F12 medium. Lipid droplets, morphology, activation markers (α-SMA, collagen-I, fibronectin), and stemness-associated markers were analysed via qRT-PCR and immunofluorescence.

Results: Lrat co-localized with Oct4/Nanog in mice. MSC-cultured PSCs had more lipid droplets, a quiescent morphology, and lower levels of activation markers than DMEM/F12-cultured cells. They also expressed stemness-associated markers (Pdx1, Oct4, Nkx6.1, Ngn3, Sox2, Pax6).

Discussion: These results indicate that PSCs possess stemness and can be preserved under specific culture conditions. The ability of MSC medium to maintain PSCs in a quiescent and stem-like state provides a valuable model for studying their biology.

Conclusion: PSCs exhibit stemness, and MSC medium maintains their quiescence, offering a novel experimental platform to study PSC biology and lineage plasticity.