Abstract
BACKGROUND: This study systematically evaluated the potential of porcine platelet-lysate (pPL) as a fetal bovine serum (FBS) substitute in cell culture. By culturing tumor cells, adult cells, and stem cells, it analyzed the differences and advantages of pPL over FBS. METHODS: pPL was prepared using the freeze - thaw method. Its biological effects were assessed by cell counting, LDH cytotoxicity assays, flow cytometry, and adipogenic/osteogenic/chondrogenic differentiation experiments, combined with qPCR and histological staining. RESULTS: pPL significantly promoted the proliferation of A375 and HDF cells, with much lower LDH release than the FBS group, indicating less cellular stress. Flow cytometry showed that MSCs in the pPL group stably expressed mesenchymal markers CD73 ⁺ /CD90 ⁺ (>95%) and had lowCD34 ⁻ (＜2％), meeting ISCT standards. Differentiation experiments revealed that the pPL group had better osteogenic differentiation efficiency, similar chondrogenic ability, and slightly lower but normal adipogenic differentiation compared to the FBS group. CONCLUSIONS: pPL effectively supports the in vitro expansion of various cells, helps maintain the immunophenotypic stability of MSCs, and shows superiority in osteogenic differentiation. With its unique "proliferation-first, differentiation-controllable" advantage from a natural growth factor network, pPL offers a new approach for developing FBS substitutes.