@article{f37f91c5f143482e94c99899a4f84fb9,
title = "A New Microengineered Platform for 4D Tracking of Single Cells in a Stem-Cell-Based In Vitro Morphogenesis Model",
abstract = "Recently developed stem-cell-based in vitro models of morphogenesis can help shed light on the mechanisms involved in embryonic patterning. These models are showcased using traditional cell culture platforms and materials, which allow limited control over the biological system and usually do not support high-content imaging. In contrast, using advanced microengineered tools can help in microscale control, long-term culture, and real-time data acquisition from such biological models and aid in elucidating the underlying mechanisms. Here, a new culturing, manipulation and analysis platform is described to study in vitro morphogenesis using thin polycarbonate film-based microdevices. A pipeline consisting of open-source software to quantify 3D cell movement using 4D image acquisition is developed to analyze cell migration within the multicellular clusters. It is shown that the platform can be used to control and study morphogenesis in non-adherent cultures of the P19C5 mouse stem cell line and mouse embryonic stem cells (mESCs) that show symmetry breaking and axial elongation events similar to early embryonic development. Using the new platform, it is found that localized cell proliferation and coordinated cell migration result in elongation morphogenesis of the P19C5 aggregates. Further, it is found that polarization and elongation of mESC aggregates are dependent on directed cell migration.",
keywords = "3D cell tracking, embryoid bodies, gastruloids, microengineering, morphogenesis, DISRUPT MICROFILAMENT ORGANIZATION, BLASTOCYST-LIKE STRUCTURES, ANTERIOR-POSTERIOR AXIS, DEVELOPMENTAL TOXICITY, LONG-TERM, GASTRULATION MODEL, ELONGATION, EMBRYOS, ESTABLISHMENT, LATRUNCULINS",
author = "Pinak Samal and Philipp Maurer and {van Blitterswijk}, Clemens and Roman Truckenmuller and Stefan Giselbrecht",
note = "Funding Information: The authors acknowledge financial support by the Dutch Province of Limburg (program {\textquoteleft}Limburg INvesteert in haar Kenniseconomie/LINK'). This research has received funding from the European Research Council (ERC) under the European Union's Horizon 2020 research and innovation programme grant agreement No 694801). The authors acknowledge financial support from Stichting De Weijerhorst. The gastruloid work was supported by an EMBO Short Term Fellowship (EMBO ASTF 78–2016) and a Company of Biologists Travelling Fellowship (DEVTF‐151202) to P.S. The authors would like to acknowledge the help from Dr. Yusuke Marikawa (University of Hawaii) for the P19 and P19C5 cells as well as help for setting up the morphogenesis model. The cells used in this study were the P19C5 cell line, which are a “clonal sub‐line” of P19 cells obtained from the American Type Culture Collection, isolated by Dr. Marikawa's group. The authors would also like to acknowledge the help received from Hang Nguyen (Maastricht University) for her critical feedback regarding the text and helpful discussions. The authors acknowledge help from Professor Alfonso Martinez‐Arias (University of Cambridge) and Tina Balayo during setting up of the gastruloid culture and for the TCF/LEF::mCherry mESCs. Funding Information: The authors acknowledge financial support by the Dutch Province of Limburg (program ?Limburg INvesteert in haar Kenniseconomie/LINK'). This research has received funding from the European Research Council (ERC) under the European Union's Horizon 2020 research and innovation programme grant agreement No 694801). The authors acknowledge financial support from Stichting De Weijerhorst. The gastruloid work was supported by an EMBO Short Term Fellowship (EMBO ASTF 78?2016) and a Company of Biologists Travelling Fellowship (DEVTF-151202) to P.S. The authors would like to acknowledge the help from Dr. Yusuke Marikawa (University of Hawaii) for the P19 and P19C5 cells as well as help for setting up the morphogenesis model. The cells used in this study were the P19C5 cell line, which are a ?clonal sub-line? of P19 cells obtained from the American Type Culture Collection, isolated by Dr. Marikawa's group. The authors would also like to acknowledge the help received from Hang Nguyen (Maastricht University) for her critical feedback regarding the text and helpful discussions. The authors acknowledge help from Professor Alfonso Martinez-Arias (University of Cambridge) and Tina Balayo during setting up of the gastruloid culture and for the TCF/LEF::mCherry mESCs. Publisher Copyright: {\textcopyright} 2020 The Authors. Published by WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim",
year = "2020",
month = jun,
doi = "10.1002/adma.201907966",
language = "English",
volume = "32",
journal = "Advanced Materials",
issn = "0935-9648",
publisher = "Wiley-VCH Verlag",
number = "24",
}