REPLACEMENT OF FETAL BOVINE SERUM BY HUMAN BLOOD SERUM: FULLY AUTOMATED SERUM PRODUCTION AND EFFECT ON FIBROBLAST CELL PROLIFERATION
Katharina Düregger, Jasmin Kreh, Ma rkus Eblenkamp
Institute of Medical and Polymer Engineering, Technical University of Munich, Germany
Growth media for in vitro cell cultures are commonly supplemented with Fetal Bovine Serum (FBS) to enhance cell proliferation. Recent discussions regarding ethical concerns towards production, as well as high variations between batches of FBS highlight the need for a replacement. Human blood serum is discussed as a possible alternative especially for autologous cell-based therapies reducing the risk of infection and immunological responses initiated by xenogeneic materials. In this study we successfully produced human blood serum from whole blood with two approaches and tested the effect on cell proliferation compared to FBS. Aiming at a high amount of growth factors and an uncomplicated production process two methods for serum production were investigated. Method 1: Platelet Concentrate (PC) was produced from citrated whole blood with a fully automated centrifugation system. Activation of platelets and release of growth factors were enhanced by a freezing and thawing cycle followed by further centrifugation. The supernatant containing the growth factors was used as FBS replacement 1. Method 2: Whole blood was dynamically incubated with glass beads resulting in activation of platelets, release of growth factors, and coagulation. Serum was then produced by removing the cellular and fibrin clot components with the same fully automated centrifugation system and used as FBS replacement 2. In both replacements 1 and 2 the amounts of PDGF and TGF-ß were measured using the ELISA method. Cell proliferation of fibroblasts cultivated with different amounts of replacement 1 and 2 as additives to the cell culture medium was compared to standard cultivation with FBS.
The results showed that stimulation of cell proliferation by human blood serum is comparable to the effect of FBS. Thus the study indicates that replacing fetal bovine serum by human blood serum is a promising strategy especially for use in autologous cell-based therapies.