Vascularized and Complex Organ Buds from Diverse Tissues via Mesenchymal Cell-Driven Condensation.

Transplantation of in-vitro-generated organ buds is a promising approach toward regenerating functional and vascularized organs. Though it has been recently shown in the context of liver models, demonstrating the applicability of this approach to other systems by delineating the molecular mechanisms guiding organ bud formation is critical. Here, we demonstrate a generalized method for organ bud formation from diverse tissues by combining pluripotent stem cell-derived tissue-specific progenitors or relevant tissue samples with endothelial cells and mesenchymal stem cells (MSCs). The MSCs initiated condensation within these heterotypic cell mixtures, which was dependent upon substrate matrix stiffness. Defining optimal mechanical properties promoted formation of 3D, transplantable organ buds from tissues including kidney, pancreas, intestine, heart, lung, and brain. Transplanted pancreatic and renal buds were rapidly vascularized and self-organized into functional, tissue-specific structures. These findings provide a general platform for harnessing mechanical properties to generate vascularized, complex organ buds with broad applications for regenerative medicine.

[1]  Takanori Takebe,et al.  Vascularized and functional human liver from an iPSC-derived organ bud transplant , 2013, Nature.

[2]  M. S. Steinberg,et al.  ON THE MECHANISM OF TISSUE RECONSTRUCTION BY DISSOCIATED CELLS, III. FREE ENERGY RELATIONS AND THE REORGANIZATION OF FUSED, HETERONOMIC TISSUE FRAGMENTS. , 1962, Proceedings of the National Academy of Sciences of the United States of America.

[3]  D. Discher,et al.  Crawling from soft to stiff matrix polarizes the cytoskeleton and phosphoregulates myosin-II heavy chain , 2012, The Journal of cell biology.

[4]  G. Forgacs,et al.  Viscoelastic properties of living embryonic tissues: a quantitative study. , 1998, Biophysical journal.

[5]  Wolfgang Huber,et al.  Directional tissue migration through a self-generated chemokine gradient , 2013, Nature.

[6]  Juergen A. Knoblich,et al.  Organogenesis in a dish: Modeling development and disease using organoid technologies , 2014, Science.

[7]  Bruce J Aronow,et al.  A catalogue of gene expression in the developing kidney. , 2003, Kidney international.

[8]  H. Haga,et al.  Diphosphorylation of the myosin regulatory light chain enhances the tension acting on stress fibers in fibroblasts , 2006, Journal of cellular physiology.

[9]  Hiroyuki Miyoshi,et al.  Self-formation of functional adenohypophysis in three-dimensional culture , 2011, Nature.

[10]  Kenneth M. Yamada,et al.  Defects in Cell Adhesion and the Visceral Endoderm following Ablation of Nonmuscle Myosin Heavy Chain II-A in Mice* , 2004, Journal of Biological Chemistry.

[11]  P. Janmey,et al.  Tissue Cells Feel and Respond to the Stiffness of Their Substrate , 2005, Science.

[12]  K. Sekine,et al.  The generation of pancreatic β-cell spheroids in a simulated microgravity culture system. , 2013, Biomaterials.

[13]  T. Svitkina,et al.  Functions of Nonmuscle Myosin II in Assembly of the Cellular Contractile System , 2012, PloS one.

[14]  Takanori Takebe,et al.  Generation of a vascularized and functional human liver from an iPSC-derived organ bud transplant , 2014, Nature Protocols.

[15]  B. Schermer,et al.  Tracking the fate of glomerular epithelial cells in vivo using serial multiphoton imaging in novel mouse models with fluorescent lineage tags , 2013, Nature Medicine.

[16]  Sami Alom Ruiz,et al.  Mechanical tugging force regulates the size of cell–cell junctions , 2010, Proceedings of the National Academy of Sciences.

[17]  S. Sen,et al.  Matrix Elasticity Directs Stem Cell Lineage Specification , 2006, Cell.

[18]  L. Sulak,et al.  Myosin-dependent junction remodelling controls planar cell intercalation and axis elongation , 2004, Nature.

[19]  Chad A. Cowan,et al.  Liver in a dish , 2013, Cell Research.

[20]  A. Brunet,et al.  Mechanical Signals Trigger Myosin II Redistribution and Mesoderm Invagination in Drosophila Embryos , 2009, Science Signaling.

[21]  Yoshiki Sasai,et al.  Cytosystems dynamics in self-organization of tissue architecture , 2013, Nature.

[22]  Dennis E Discher,et al.  How deeply cells feel: methods for thin gels , 2010, Journal of physics. Condensed matter : an Institute of Physics journal.

[23]  K. Sekine,et al.  Self-organization of human hepatic organoid by recapitulating organogenesis in vitro. , 2012, Transplantation proceedings.

[24]  Asako Shindo,et al.  PCP and Septins Compartmentalize Cortical Actomyosin to Direct Collective Cell Movement , 2014, Science.

[25]  J. Rossant,et al.  Liver Organogenesis Promoted by Endothelial Cells Prior to Vascular Function , 2001, Science.

[26]  H. Clevers,et al.  Growing Self-Organizing Mini-Guts from a Single Intestinal Stem Cell: Mechanism and Applications , 2013, Science.

[27]  W. Kern Cleaning solutions based on hydrogen peroxide for use in silicon semiconductor technology , 1970 .

[28]  Hiromu Suzuki,et al.  Transient vascularization of transplanted human adult-derived progenitors promotes self-organizing cartilage. , 2014, The Journal of clinical investigation.

[29]  U. S. Schwarz,et al.  Coupling biochemistry and mechanics in cell adhesion: a model for inhomogeneous stress fiber contraction , 2007, 0707.2551.

[30]  P. Greer,et al.  Reduced Cell Migration and Disruption of the Actin Cytoskeleton in Calpain-deficient Embryonic Fibroblasts* , 2001, The Journal of Biological Chemistry.

[31]  Ondine Cleaver,et al.  Induction of Pancreatic Differentiation by Signals from Blood Vessels , 2001, Science.

[32]  M. Takeichi Self-organization of animal tissues: cadherin-mediated processes. , 2011, Developmental cell.

[33]  T. Adachi,et al.  Self-organizing optic-cup morphogenesis in three-dimensional culture , 2011, Nature.

[34]  D. Discher,et al.  Optimal matrix rigidity for stress fiber polarization in stem cells. , 2010, Nature physics.

[35]  Peter Friedl,et al.  Compensation mechanism in tumor cell migration , 2003, The Journal of cell biology.

[36]  Timothy J Mitchison,et al.  Dissecting Temporal and Spatial Control of Cytokinesis with a Myosin II Inhibitor , 2003, Science.

[37]  C. Hunter,et al.  Contractile forces sustain and polarize hematopoiesis from stem and progenitor cells. , 2014, Cell stem cell.

[38]  Y. Sasai Next-generation regenerative medicine: organogenesis from stem cells in 3D culture. , 2013, Cell stem cell.