ESTABLISHMENT OF A LYMPHOBLAST CULTURE PROTOCOL FROM PERIPHERAL BLOOD

Tóm tắt

Lymphoblasts, as lymphocyte precursors, are vital for research in cellular biology, immunology, genetics, and therapeutic development due to their proliferative potential. This study introduces a standardized protocol for isolating, transforming, and culturing lymphoblasts from peripheral blood of three healthy volunteers (≥18 years). The method uses Ficoll-Hypaque gradient centrifugation to isolate mononuclear cells, followed by Epstein-Barr virus (EBV) transformation and culture in RPMI-1640 medium with fetal bovine serum (FBS), phytohemagglutinin (PHA), and antibiotics at 37°C with 5% CO₂. Growth was assessed via Giemsa staining, automated cell counting, and Trypan Blue viability assays. The protocol achieved high lymphocyte purity (up to 90%), a mean 37-fold proliferation over 12 days, and 90.5% viability, ensuring reproducibility across samples. This standardized approach enables reproducible high-density lymphoblast cultures from minimal blood volumes, offering a robust platform for advancing studies in immunology, genetics, metabolic disorders, and cancer vaccine development. These results support progress in biotechnology research and therapeutic innovation.