BHLHE40 Maintains the Stemness of PαS Cells In Vitro by Targeting Zbp1 through the Wnt/β-Catenin Signaling Pathway

Primary bone mesenchymal stem cells (BMSCs) gradually lose stemness during in vitro expansion, which significantly affects the cell therapeutic effects. Here, we chose murine PαS (SCA-1+PDGFRα+CD45−TER119−) as representative of BMSCs and aimed to explore premium culture conditions for cells. Freshly isolated (fresh) were obtained from limbs C57/6N mice by fluorescence-activated sorting (FACS). We investigated differences proliferation, differentiation, markers ectopic osteogenesis chondrogenesis ability vivo, well changes expansion vitro. Gain- loss-of-function experiments applied investigate critical role underlying mechanism basic helix–loop–helix family member E40 (BHLHE40) maintaining The fresh vivo was superior that passage 0 (P0) decreased P0 4 (P4). Moreover, BHLHE40 plays a regulating expansion. Mechanically, regulates targeting Zbp1 through Wnt/β-catenin signaling pathway. This work confirms is factor may provide significant indications exploration