reproducible, ultra high-throughput formation of multicellular organization from single cell suspension-derived human embryonic stem cell aggregates可再生的、超高效的形成多细胞组织单细胞suspension-derived人类胚胎干细胞聚集.pdfVIP

Reproducible, Ultra High-Throughput Formation of Multicellular Organization from Single Cell Suspension- Derived Human Embryonic Stem Cell Aggregates 1 1 1 ´ 1,2 1,2 Mark D. Ungrin , Chirag Joshi , Andra Nica , Celine Bauwens , Peter W. Zandstra * 1 Institute of Biomaterials and Biomedical Engineering, University of Toronto, Toronto, Ontario, Canada, 2 Department of Chemical Engineering and Applied Chemistry, University of Toronto, Toronto, Ontario, Canada Abstract Background: Human embryonic stem cells (hESC) should enable novel insights into early human development and provide a renewable source of cells for regenerative medicine. However, because the three-dimensional hESC aggregates [embryoid bodies (hEB)] typically employed to reveal hESC developmental potential are heterogeneous and exhibit disorganized differentiation, progress in hESC technology development has been hindered. Methodology/Principal Findings: Using a centrifugal forced-aggregation strategy in combination with a novel centrifugal- extraction approach as a foundation, we demonstrated that hESC input composition and inductive environment could be manipulated to form large numbers of well-defined aggregates exhibiting multi-lineage differentiation and substantially improved self-organization from single-cell suspensions. These aggregates exhibited coordinated bi-domain structures including contiguous regions of extraembryonic endoderm- and epiblast-like tissue. A silicon wafer-based microfabrication technology was used to generate surfaces that permit the production of hundreds to thousands of hEB per cm2. Conclusions/Significance: The mechanisms of early human embryogenesis are poorly understood. We report an ultra