@article{ee6c8bd272e34c3aacfd1d952af8a871,
title = "Protocol for differentiation of bovine adipogenic progenitor cells embedded in alginate sheets",
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.",
keywords = "Cell Biology, Cell Culture, Cell Differentiation, Cell Isolation, Cell-based Assays, High-throughput Screening, Microscopy, Stem Cells",
author = "Caponi, {Joseph J.} and Jane Shmushkis and Hanna Geissler and Post, {Mark J.} and Laura Jackisch and Mall, {Eva M.}",
note = "Funding Information: J.J.C., J.S., H.G., L.J., and E.M.M are employees of Mosa Meat B.V. M.J.P. is a co-founder and stakeholder of Mosa Meat B.V. This study was funded by Mosa Meat B.V. Mosa Meat B.V. has patents pending on the use of FAPs for cultured fat production (JBB/P129698NL00) and serum-free differentiation medium for adipogenesis (JBB/P129695NL00). J.S. was employed by Opentrons (New York, New York) from Feb 2018 to Aug 2020. Funding Information: The work was funded by Mosa Meat B.V. We would like to acknowledge Cheyenne Hendriks, Dirk Remmers, and Christoph B{\"o}rlin (Computing Department) for assistance with image acquisition and data analysis. All members of the Fat Tissue Engineering department helped with cell preparations and provided valuable feedback on experimental design, protocol optimizations, and data interpretation. Special thanks go to Federica Cantoni for excellent technical assistance and Sebastiano Alberganti for initial tests on coatings that help attach alginate mixtures to the well plate. Eric Wieland (Automation Team) provided valuable input on the initial protocol design, and Sander Purnot (Automation Team) provided help with the design of customized labware solutions. J.J.C. and E.M.M. conceptualized and developed the presented protocol. J.J.C. H.G. and J.S. wrote and optimized the Python code for the liquid handler. E.M.M. performed cell culture experiments and data analysis. M.J.P. L.J. and E.M.M. supervised the study and took part in scientific discussions of the experimental data. J.J.C. and E.M.M. wrote the manuscript with input from all authors. J.J.C. J.S. H.G. L.J. and E.M.M are employees of Mosa Meat B.V. M.J.P. is a co-founder and stakeholder of Mosa Meat B.V. This study was funded by Mosa Meat B.V. Mosa Meat B.V. has patents pending on the use of FAPs for cultured fat production (JBB/P129698NL00) and serum-free differentiation medium for adipogenesis (JBB/P129695NL00). J.S. was employed by Opentrons (New York, New York) from Feb 2018 to Aug 2020. Funding Information: The work was funded by Mosa Meat B.V. We would like to acknowledge Cheyenne Hendriks, Dirk Remmers, and Christoph B{\"o}rlin (Computing Department) for assistance with image acquisition and data analysis. All members of the Fat Tissue Engineering department helped with cell preparations and provided valuable feedback on experimental design, protocol optimizations, and data interpretation. Special thanks go to Federica Cantoni for excellent technical assistance and Sebastiano Alberganti for initial tests on coatings that help attach alginate mixtures to the well plate. Eric Wieland (Automation Team) provided valuable input on the initial protocol design, and Sander Purnot (Automation Team) provided help with the design of customized labware solutions. Publisher Copyright: {\textcopyright} 2023 The Author(s)",
year = "2023",
month = jun,
day = "16",
doi = "10.1016/j.xpro.2023.102143",
language = "English",
volume = "4",
journal = "STAR protocols",
issn = "2666-1667",
publisher = "Elsevier",
number = "2",
}