Evaluation of human Platelet Lysate in Animal-Free Conditions for Cell Culture and Cytotoxicity Testing Alexander Edwards 1 , Christopher Longmore 1 , Lottie Roscoe 1 , Carol Treasure 1 . 1 XCELLR8 Ltd, The Innovation Centre, Sci-Tech Daresbury, Cheshire WA4 4FS, UK Human Platelet Lysate (hPL) is commonly used as an animal-free alternative to FBS/FCS. Currently, Human Serum is used for animal-free cell culture conditions at XCellR8, as we aim to replace the use of animal derived products in the laboratory. Human Platelet Lysate is comparable on price to Human Serum. The aim of this work was to investigate the use of hPL for animal-free cell culture systems in comparison with Human Serum for both cell growth, viability and functional cytotoxicity testing. Methods Human Platelet Lysate supported comparable levels of TK6 and Fibroblast cell proliferation and cell viability at all test concentrations compared with Human Serum. Cell morphology was unchanged between test and control conditions for both TK6 (not shown) and Fibroblast cells. The results for functional cytotoxicity testing using TK6 showed that 5% and 2.5% hPL produced similar performance compared with 10% human serum. For Fibroblasts, 2% hPL gave the most comparable results compared with 2% HS. In conclusion, hPL offers a viable and cost effective alternative to HS for cell culture models. RESULTS *p<0.05 vs. 10% HiHs (t-test). n=6. Data from 2 independent experiments. *p<0.05 vs. 10% HiHs (t-test). Data from 2 independent experiments shown; n=3 per experiment. *p<0.05 vs. 10% HiHs (t-test). PC data from 2 independent experiments shown; n=12 per experiment. One representative experiment for NC shown, not significant in 2 independent experiments. *p<0.05 vs. 10% HiHs (t-test). n=6. Data from 2 independent experiments. *p<0.05 vs. 10% HS (t-test). Data from 2 independent experiments shown; n=3 per experiment. *p<0.05 vs. 10% HS (t-test). PC data from 2 independent experiments shown; n=12 per experiment. One representative experiment for NC shown, not significant in 2 independent experiments. Gluc-TK6 (Lymphoblastoid) HDFn-XF (Fibroblast) *p<0.05 vs. 10% HS (t-test). n=6. Data from 2 independent experiments. *p<0.05 vs. 10% HS (t-test). n=6. Data from 2 independent experiments. Morphology of Fibroblasts over a time course: Human Serum (2%) vs hPL (2, 1, 0.5%). INTRODUCTION / AIM Cell Growth Cell Viability Functional Cytotoxicity Testing Functional Cytotoxicity Testing Cell Growth Cell Viability Morphology DISCUSSION