Abstract
Here, we present a cost-effective protocol to differentiate bovine fibro-adipogenic progenitors in a thin hydrogel sheet adherent to 96-well plates. We describe steps for the embedding and culturing of cells in alginate sheets, culture maintenance, and analysis. Compared to alternative three-dimensional (3D) models such as hydrogel-based microfibers, this approach simplifies automation while retaining efficient maturation of adipocytes. Embedded cells are still subjected to a 3D environment, but the sheets can be handled and analyzed like two-dimensional cultures.