Low-dose dopamine reduces inflammatory factors of acute pancreatitis in rats.

The optimization of culture conditions for primary vascular cells remains challenging, as cell viability alone does not necessarily reflect preserved molecular identity or functional stability. Human umbilical vein endothelial cells (HUVECs) and Human aortic vascular smooth muscle cells (T/G HA-VSMCs) are highly sensitive to their microenvironment, and non-specific or poorly standardized supplements may sustain survival while disrupting gene expression and phenotypic integrity. Human platelet lysate (HPL) has emerged as a human-derived alternative to conventional supplements. However, its role as a supplement rather than a complete serum replacement remains insufficiently explored in vascular cell culture systems. This research innovates by evaluating HPL and CM as complementary supplements to FBS rather than standard specific medium. HUVECs and T/G HA-VSMC were cultured in Dulbecco's Modified Eagle Medium supplemented with 10% fetal bovine serum (FBS) and either 5% or 10% HPL or fibroblast-derived conditioned medium (CM). Cell viability was assessed using MTT assay, morphological characteristics were evaluated by phase-contrast microscopy, and lineage-specific gene expression was analyzed by quantitative real-time PCR targeting endothelial (CD31, CD144) and smooth muscle (ACTA2, SM22α) markers. Platelet lysates derived from both fresh and expired platelet concentrates were evaluated. HPL supplementation significantly enhanced cell viability and preserved characteristic morphology in both cell types compared to CM. Importantly, no significant alterations in lineage-specific gene expression were observed following HPL supplementation, indicating maintenance of cellular identity. Comparable biological outcomes were obtained using HPL prepared from fresh and expired platelet concentrates, suggesting retained growth factor activity beyond clinical transfusion applicability. Human platelet lysate, when used as a supplement to fetal bovine serum, supports the growth and viability of human umbilical vein endothelial cells and vascular smooth muscle cells while preserving the expression of key phenotypic markers. Compared to conditioned media, HPL demonstrated more stable effects on gene expression and cell morphology. Additionally, the use of expired platelet concentrates provides a cost-effective and practical approach for cell culture applications.