(91d) Development of an Animal-Free Endothelial Cell Culture Media and Adaptation Method

The successful development of a defined media for cell culture presents a host of advantages including reduction of disease transmission, facilitation of translational research, and enablement of finely tuned bioassays. Fetal bovine serum (FBS) has a nearly universal use in cell culture, but it presents consequential drawbacks, such as significant variability from batch-to-batch, serious risk of disease transmission and concerns regarding ethical collection practices. As interest continues to grow around the development of biomimetic in vitro assays and translational biomedical research, the need for full identification of components in cell media and stimulating factors have become increasingly important. Here, we present our formulation for an animal-free (AF) media and our adaptation procedures for human umbilical vein endothelial cell (HUVEC). Our goal in this study is to identify the simplest media formulation that is capable of sustaining viable HUVEC growth while optimizing for the most efficient route to adapting cells to AF conditions. This effort is intended to preserve a sufficient number of fully AF passages for experimentation. We found that coating the culture vessel with attachment proteins is critical under conditions of high or fully AF media to enhance attachment of the seeded cells and improve proliferation. We obtained conditions of cell culture that were fully AF in a chemically-defined media. This work outlines the methodologies and reasoning for implementing specific steps during the adaptation process that can be used for HUVECs and other human cell lines, and it can motivate the development of quantitative, tunable bioassays to accelerate biomedical research and translation.