Disease modelling in human organoids

ABSTRACT The past decade has seen an explosion in the field of in vitro disease modelling, in particular the development of organoids. These self-organizing tissues derived from stem cells provide a unique system to examine mechanisms ranging from organ development to homeostasis and disease. Because organoids develop according to intrinsic developmental programmes, the resultant tissue morphology recapitulates organ architecture with remarkable fidelity. Furthermore, the fact that these tissues can be derived from human progenitors allows for the study of uniquely human processes and disorders. This article and accompanying poster highlight the currently available methods, particularly those aimed at modelling human biology, and provide an overview of their capabilities and limitations. We also speculate on possible future technological advances that have the potential for great strides in both disease modelling and future regenerative strategies. Summary: Human organoids are important tools for modelling disease. This At a Glance article summarises the current organoid models of several human diseases, and discusses future prospects for these technologies.

[1]  A. Anthony,et al.  Growth in culture of trypsin dissociated thyroid cells from adult rats. , 1966, Experimental cell research.

[2]  George E. Allen,et al.  Human embryonic lung epithelial tips are multipotent progenitors that can be expanded in vitro as long-term self-renewing organoids , 2017, eLife.

[3]  Hans Clevers,et al.  Tissue-specific mutation accumulation in human adult stem cells during life , 2016, Nature.

[4]  B. Stripp,et al.  Lung Stem Cell Differentiation in Mice Directed by Endothelial Cells via a BMP4-NFATc1-Thrombospondin-1 Axis , 2014, Cell.

[5]  Cyriac Kandoth,et al.  Tumor Evolution and Drug Response in Patient-Derived Organoid Models of Bladder Cancer , 2018, Cell.

[6]  B. Göttgens,et al.  Lgr5+ stem and progenitor cells reside at the apex of a heterogeneous embryonic hepatoblast pool , 2019, Development.

[7]  M J Bissell,et al.  Lumen formation by epithelial cell lines in response to collagen overlay: a morphogenetic model in culture. , 1982, Proceedings of the National Academy of Sciences of the United States of America.

[8]  T. Karlsen,et al.  Reconstruction of the mouse extrahepatic biliary tree using primary human extrahepatic cholangiocyte organoids , 2017, Nature Medicine.

[9]  Davide Prandi,et al.  Personalized In Vitro and In Vivo Cancer Models to Guide Precision Medicine. , 2017, Cancer discovery.

[10]  Manuel Hidalgo,et al.  Patient-derived xenograft models: an emerging platform for translational cancer research. , 2014, Cancer discovery.

[11]  H. Baharvand,et al.  Efficient Differentiation of Human Embryonic Stem Cells Toward Dopaminergic Neurons Using Recombinant LMX1A Factor , 2015, Molecular Biotechnology.

[12]  D. Tuveson,et al.  Successful creation of pancreatic cancer organoids by means of EUS-guided fine-needle biopsy sampling for personalized cancer treatment. , 2017, Gastrointestinal endoscopy.

[13]  Qisheng Zhang,et al.  High-Content Screening in hPSC-Neural Progenitors Identifies Drug Candidates that Inhibit Zika Virus Infection in Fetal-like Organoids and Adult Brain. , 2017, Cell stem cell.

[14]  Hans Clevers,et al.  Long-term expansion of epithelial organoids from human colon, adenoma, adenocarcinoma, and Barrett's epithelium. , 2011, Gastroenterology.

[15]  M. Belletrutti,et al.  A “living” will , 2008, Canadian Medical Association Journal.

[16]  Michael Schumacher,et al.  Modeling human development and disease in pluripotent stem cell-derived gastric organoids , 2014, Nature.

[17]  Hongmei Mou,et al.  Dual SMAD Signaling Inhibition Enables Long-Term Expansion of Diverse Epithelial Basal Cells. , 2016, Cell stem cell.

[18]  S. Janes,et al.  The secret lives of cancer cell lines , 2018, Disease Models & Mechanisms.

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

[20]  M. Zernicka-Goetz,et al.  From pluripotency to differentiation: laying foundations for the body pattern in the mouse embryo , 2014, Philosophical Transactions of the Royal Society B: Biological Sciences.

[21]  Harry Begthel,et al.  Mouse and human urothelial cancer organoids: A tool for bladder cancer research , 2019, Proceedings of the National Academy of Sciences.

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

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

[24]  D. Graham,et al.  Replication of human noroviruses in stem cell–derived human enteroids , 2016, Science.

[25]  T. Teshima,et al.  Overview of Human Salivary Glands: Highlights of Morphology and Developing Processes , 2017, Anatomical record.

[26]  Hans Clevers,et al.  Unlimited in vitro expansion of adult bi-potent pancreas progenitors through the Lgr5/R-spondin axis , 2013, The EMBO journal.

[27]  Markus H. Heim,et al.  Organoid Models of Human Liver Cancers Derived from Tumor Needle Biopsies , 2018, Cell reports.

[28]  E. Cuppen,et al.  Generation and characterization of rat liver stem cell lines and their engraftment in a rat model of liver failure , 2016, Scientific Reports.

[29]  Yoshiki Sasai,et al.  Self-formation of optic cups and storable stratified neural retina from human ESCs. , 2012, Cell stem cell.

[30]  T. Karlsen,et al.  Cholangiocytes derived from human induced pluripotent stem cells for disease modeling and drug validation , 2015, Nature Biotechnology.

[31]  D. Stolz,et al.  Histological organization in hepatocyte organoid cultures. , 2001, The American journal of pathology.

[32]  K. Mizuseki,et al.  Directed differentiation of telencephalic precursors from embryonic stem cells , 2005, Nature Neuroscience.

[33]  Fabian J Theis,et al.  Quantification of regenerative potential in primary human mammary epithelial cells , 2015, Development.

[34]  J. Spence,et al.  Neural Crest Cell Implantation Restores Enteric Nervous System Function and Alters the Gastrointestinal Transcriptome in Human Tissue-Engineered Small Intestine , 2017, Stem cell reports.

[35]  Hans Clevers,et al.  Long-Term Culture of Genome-Stable Bipotent Stem Cells from Adult Human Liver , 2015, Cell.

[36]  R. Shivdasani,et al.  Co-culture of Gastric Organoids and Immortalized Stomach Mesenchymal Cells. , 2016, Methods in molecular biology.

[37]  Hans Clevers,et al.  Functional repair of CFTR by CRISPR/Cas9 in intestinal stem cell organoids of cystic fibrosis patients. , 2013, Cell stem cell.

[38]  S. O’Rahilly,et al.  Trophoblast organoids as a model for maternal–fetal interactions during human placentation , 2018, Nature.

[39]  A. Ranga,et al.  Artificial three-dimensional niches deconstruct pancreas development in vitro , 2013, Development.

[40]  J. Bagley,et al.  Genetically engineered cerebral organoids model brain tumour formation , 2018, Nature Methods.

[41]  M. Long,et al.  Three-dimensional cellular development is essential for ex vivo formation of human bone , 2000, Nature Biotechnology.

[42]  L. Hayflick THE LIMITED IN VITRO LIFETIME OF HUMAN DIPLOID CELL STRAINS. , 1965, Experimental cell research.

[43]  Ping Liu,et al.  25‐Hydroxycholesterol Protects Host against Zika Virus Infection and Its Associated Microcephaly in a Mouse Model , 2017, Immunity.

[44]  Hans C Clevers,et al.  Rectal Organoids Enable Personalized Treatment of Cystic Fibrosis. , 2019, Cell reports.

[45]  A. van Oudenaarden,et al.  Expansion of Adult Human Pancreatic Tissue Yields Organoids Harboring Progenitor Cells with Endocrine Differentiation Potential , 2018, Stem cell reports.

[46]  F. Müller,et al.  An Organoid-Based Model of Cortical Development Identifies Non-Cell-Autonomous Defects in Wnt Signaling Contributing to Miller-Dieker Syndrome. , 2017, Cell reports.

[47]  Hayley E. Francies,et al.  Organoid cultures recapitulate esophageal adenocarcinoma heterogeneity providing a model for clonality studies and precision therapeutics , 2018, Nature Communications.

[48]  D. Ingber,et al.  From 3D cell culture to organs-on-chips. , 2011, Trends in cell biology.

[49]  Hans Clevers,et al.  Sequential cancer mutations in cultured human intestinal stem cells , 2015, Nature.

[50]  Hans Clevers,et al.  A functional CFTR assay using primary cystic fibrosis intestinal organoids , 2013, Nature Medicine.

[51]  Hans Clevers,et al.  A Living Biobank of Breast Cancer Organoids Captures Disease Heterogeneity , 2018, Cell.

[52]  M. Gerstein,et al.  FOXG1-Dependent Dysregulation of GABA/Glutamate Neuron Differentiation in Autism Spectrum Disorders , 2015, Cell.

[53]  Takanori Kanai,et al.  Modeling colorectal cancer using CRISPR-Cas9–mediated engineering of human intestinal organoids , 2015, Nature Medicine.

[54]  K. Sekine,et al.  Massive and Reproducible Production of Liver Buds Entirely from Human Pluripotent Stem Cells. , 2017, Cell reports.

[55]  M J Bissell,et al.  Influence of a reconstituted basement membrane and its components on casein gene expression and secretion in mouse mammary epithelial cells. , 1987, Proceedings of the National Academy of Sciences of the United States of America.

[56]  Alex A. Pollen,et al.  Human iPSC-Derived Cerebral Organoids Model Cellular Features of Lissencephaly and Reveal Prolonged Mitosis of Outer Radial Glia. , 2017, Cell stem cell.

[57]  Jay Shendure,et al.  The haplotype-resolved genome and epigenome of the aneuploid HeLa cancer cell line , 2013, Nature.

[58]  R. Nishinakamura,et al.  Redefining the in vivo origin of metanephric nephron progenitors enables generation of complex kidney structures from pluripotent stem cells. , 2014, Cell stem cell.

[59]  H. Clevers,et al.  Modeling Cryptosporidium infection in human small intestinal and lung organoids , 2018, Nature Microbiology.

[60]  Dirk Timmerman,et al.  Development of organoids from mouse and human endometrium showing endometrial epithelium physiology and long-term expandability , 2017, Development.

[61]  Daniel R. Berger,et al.  Cell diversity and network dynamics in photosensitive human brain organoids , 2017, Nature.

[62]  Calvin J Kuo,et al.  Sustained in vitro intestinal epithelial culture within a Wnt-dependent stem cell niche , 2009, Nature Medicine.

[63]  Hayley E. Francies,et al.  Prospective Derivation of a Living Organoid Biobank of Colorectal Cancer Patients , 2015, Cell.

[64]  Bon-Kyoung Koo,et al.  Long-term, hormone-responsive organoid cultures of human endometrium in a chemically-defined medium , 2017, Nature Cell Biology.

[65]  Y. Chan,et al.  Directed Differentiation of Human Induced Pluripotent Stem Cells to Sensory Neurons by Combined Small Molecule Inhibitors , 2014 .

[66]  Jeffry D. Sander,et al.  CRISPR-Cas systems for editing, regulating and targeting genomes , 2014, Nature Biotechnology.

[67]  J. Wells,et al.  Wnt/β-catenin promotes gastric fundus specification in mice and humans , 2017, Nature.

[68]  J. Bagley,et al.  Fused dorsal-ventral cerebral organoids model complex interactions between diverse brain regions , 2017, Nature methods.

[69]  M. Hoffman,et al.  Salivary gland development: a template for regeneration. , 2014, Seminars in cell & developmental biology.

[70]  Takanori Takebe,et al.  Vascularized and Complex Organ Buds from Diverse Tissues via Mesenchymal Cell-Driven Condensation. , 2015, Cell stem cell.

[71]  Hideshi Kawakami,et al.  Self-organization of polarized cerebellar tissue in 3D culture of human pluripotent stem cells. , 2015, Cell reports.

[72]  Hans Clevers,et al.  Lgr5(+ve) stem cells drive self-renewal in the stomach and build long-lived gastric units in vitro. , 2010, Cell stem cell.

[73]  C. Rice,et al.  Long-Term Expansion of Functional Mouse and Human Hepatocytes as 3D Organoids , 2018, Cell.

[74]  T. Arndt,et al.  In vivo , 2019, Springer Reference Medizin.

[75]  M. Eiraku,et al.  Self-organization of axial polarity, inside-out layer pattern, and species-specific progenitor dynamics in human ES cell–derived neocortex , 2013, Proceedings of the National Academy of Sciences.

[76]  David W. Nauen,et al.  Brain-Region-Specific Organoids Using Mini-bioreactors for Modeling ZIKV Exposure , 2016, Cell.

[77]  M. Asashima,et al.  Generation of stomach tissue from mouse embryonic stem cells , 2015, Nature Cell Biology.

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

[79]  H. Mollenkopf,et al.  The Notch and Wnt pathways regulate stemness and differentiation in human fallopian tube organoids , 2015, Nature Communications.

[80]  Carolina Q. Sacramento,et al.  The clinically approved antiviral drug sofosbuvir inhibits Zika virus replication , 2017, Scientific Reports.

[81]  D. Geschwind,et al.  Functional cortical neurons and astrocytes from human pluripotent stem cells in 3D culture , 2015, Nature Methods.

[82]  Hao Li,et al.  An in vivo model of functional and vascularized human brain organoids , 2018, Nature Biotechnology.

[83]  Amadou A. Sall,et al.  The Brazilian Zika virus strain causes birth defects in experimental models , 2016, Nature.

[84]  Scott H. Randell,et al.  Basal cells as stem cells of the mouse trachea and human airway epithelium , 2009, Proceedings of the National Academy of Sciences.

[85]  S. Lopes,et al.  Kidney organoids from human iPS cells contain multiple lineages and model human nephrogenesis , 2015, Nature.

[86]  L. Vallier,et al.  Directed differentiation of human induced pluripotent stem cells into functional cholangiocyte-like cells , 2017, Nature Protocols.

[87]  S. Ramaswamy,et al.  Systematic identification of genomic markers of drug sensitivity in cancer cells , 2012, Nature.

[88]  S. Akira,et al.  FGF7 is a functional niche signal required for stimulation of adult liver progenitor cells that support liver regeneration. , 2013, Genes & development.

[89]  Qiulian Wu,et al.  A Three-Dimensional Organoid Culture System Derived from Human Glioblastomas Recapitulates the Hypoxic Gradients and Cancer Stem Cell Heterogeneity of Tumors Found In Vivo. , 2016, Cancer research.

[90]  K. Tsuchihashi,et al.  Development of a functional thyroid model based on an organoid culture system. , 2018, Biochemical and biophysical research communications.

[91]  J. R. Holt,et al.  Generation of inner ear organoids with functional hair cells from human pluripotent stem cells , 2017, Nature Biotechnology.

[92]  Bryan W. Jones,et al.  Photoreceptor Outer Segment-like Structures in Long-Term 3D Retinas from Human Pluripotent Stem Cells , 2017, Scientific Reports.

[93]  Mina J Bissell,et al.  Isolation, immortalization, and characterization of a human breast epithelial cell line with stem cell properties. , 2002, Genes & development.

[94]  Daniel R Weinberger,et al.  Midbrain-like Organoids from Human Pluripotent Stem Cells Contain Functional Dopaminergic and Neuromelanin-Producing Neurons. , 2016, Cell stem cell.

[95]  Hans Clevers,et al.  Isolation and in vitro expansion of human colonic stem cells , 2011, Nature Medicine.

[96]  Jonathan A. Bernstein,et al.  Assembly of functionally integrated human forebrain spheroids , 2017, Nature.

[97]  Ruili Huang,et al.  Identification of small-molecule inhibitors of Zika virus infection and induced neural cell death via a drug repurposing screen , 2016, Nature Medicine.

[98]  Joshua M. Dempster,et al.  Genetic and transcriptional evolution alters cancer cell line drug response , 2018, Nature.

[99]  E. Cuppen,et al.  Identification of Multipotent Luminal Progenitor Cells in Human Prostate Organoid Cultures , 2014, Cell.

[100]  E. Voest,et al.  Tumor Organoids as a Pre-clinical Cancer Model for Drug Discovery. , 2017, Cell chemical biology.

[101]  Mohammad Wahid Ansari,et al.  The legal status of in vitro embryos , 2014 .

[102]  D A Scudiero,et al.  Feasibility of drug screening with panels of human tumor cell lines using a microculture tetrazolium assay. , 1988, Cancer research.

[103]  F. J. Livesey,et al.  Guided self-organization and cortical plate formation in human brain organoids , 2017, Nature Biotechnology.

[104]  Mechthild Krause,et al.  Human gastric cancer modelling using organoids , 2018, Gut.

[105]  H. Clevers,et al.  Differentiated Troy + Chief Cells Act as Reserve Stem Cells to Generate All Lineages of the Stomach Epithelium , 2013, Cell.

[106]  J. Rossant,et al.  Regeneration of Thyroid Function by Transplantation of Differentiated Pluripotent Stem Cells. , 2015, Cell stem cell.

[107]  R. Locksley,et al.  Recruited Monocytes and Type 2 Immunity Promote Lung Regeneration following Pneumonectomy. , 2017, Cell stem cell.

[108]  Mazhar Adli,et al.  The CRISPR tool kit for genome editing and beyond , 2018, Nature Communications.

[109]  Hans Clevers,et al.  Modeling Development and Disease with Organoids , 2016, Cell.

[110]  Bon-Kyoung Koo,et al.  Modeling mouse and human development using organoid cultures , 2015, Development.

[111]  F. Watt,et al.  Lineage Tracing , 2012, Cell.

[112]  Yoshiki Sasai,et al.  Generation of functional hippocampal neurons from self-organizing human embryonic stem cell-derived dorsomedial telencephalic tissue , 2015, Nature Communications.

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

[114]  Gordon Keller,et al.  Development of definitive endoderm from embryonic stem cells in culture , 2004, Development.

[115]  Gail H Deutsch,et al.  In vitro generation of human pluripotent stem cell derived lung organoids , 2015, eLife.

[116]  Michael Kyba,et al.  Generation of functional thyroid from embryonic stem cells , 2012, Nature.

[117]  M. Bjerknes,et al.  Intestinal epithelial stem cells and progenitors. , 2006, Methods in enzymology.

[118]  Laura M. Palermo,et al.  A three-dimensional model of human lung development and disease from pluripotent stem cells , 2017, Nature Cell Biology.

[119]  Orly Reiner,et al.  Human Brain Organoids on a Chip Reveal the Physics of Folding , 2018, Nature physics.

[120]  Peter M. G. Munro,et al.  Identification and Correction of Mechanisms Underlying Inherited Blindness in Human iPSC-Derived Optic Cups , 2016, Cell stem cell.

[121]  H. Clevers,et al.  Probing the Tumor Suppressor Function of BAP1 in CRISPR-Engineered Human Liver Organoids. , 2019, Cell stem cell.

[122]  O. Witte,et al.  Self‐Renewal and Multilineage Differentiation In Vitro from Murine Prostate Stem Cells , 2007, Stem cells.

[123]  M. Spector,et al.  Organoid Models of Human and Mouse Ductal Pancreatic Cancer , 2015, Cell.

[124]  Aaron M Zorn,et al.  Vertebrate endoderm development and organ formation. , 2009, Annual review of cell and developmental biology.

[125]  Olivier Gevaert,et al.  Oncogenic transformation of diverse gastrointestinal tissues in primary organoid culture , 2014, Nature Medicine.

[126]  Madeline A. Lancaster,et al.  Cerebral organoids model human brain development and microcephaly , 2013, Nature.

[127]  Bon-Kyoung Koo,et al.  Human Primary Liver Cancer -derived Organoid Cultures for disease modelling and drug screening , 2017, Nature Medicine.

[128]  M. Li,et al.  Modeling liver cancer and therapy responsiveness using organoids derived from primary mouse liver tumors , 2018, Carcinogenesis.

[129]  Michael J. Cronce,et al.  Type 2 alveolar cells are stem cells in adult lung. , 2013, The Journal of clinical investigation.

[130]  Tamas L. Horvath,et al.  Modeling human cortical development in vitro using induced pluripotent stem cells , 2012, Proceedings of the National Academy of Sciences.

[131]  G. Govindaiah,et al.  Fusion of Regionally Specified hPSC-Derived Organoids Models Human Brain Development and Interneuron Migration. , 2017, Cell stem cell.

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

[133]  Robert G. Parton,et al.  Kidney organoids from human iPS cells contain multiple lineages and model human nephrogenesis , 2016, Nature.

[134]  J. Rossant,et al.  Directed differentiation of human pluripotent stem cells into mature airway epithelia expressing functional CFTR protein , 2012, Nature Biotechnology.

[135]  Alyssa J. Miller,et al.  A bioengineered niche promotes in vivo engraftment and maturation of pluripotent stem cell derived human lung organoids , 2016, eLife.

[136]  Hans Clevers,et al.  CRISPR/Cas 9 genome editing and its applications in organoids. , 2017, American journal of physiology. Gastrointestinal and liver physiology.

[137]  Hans Clevers,et al.  A Comprehensive Human Gastric Cancer Organoid Biobank Captures Tumor Subtype Heterogeneity and Enables Therapeutic Screening. , 2018, Cell stem cell.

[138]  A. Tucker,et al.  Lineage tracing of the endoderm during oral development , 2012, Developmental dynamics : an official publication of the American Association of Anatomists.

[139]  E. Kroon,et al.  Efficient differentiation of human embryonic stem cells to definitive endoderm , 2005, Nature Biotechnology.

[140]  K. To,et al.  Differentiated human airway organoids to assess infectivity of emerging influenza virus , 2018, Proceedings of the National Academy of Sciences.

[141]  H. Clevers,et al.  Single Lgr5 stem cells build crypt–villus structures in vitro without a mesenchymal niche , 2009, Nature.

[142]  A. Hoischen,et al.  Chromosomal abnormalities in hepatic cysts point to novel polycystic liver disease genes , 2016, European Journal of Human Genetics.

[143]  Andrea Sottoriva,et al.  Patient-derived organoids model treatment response of metastatic gastrointestinal cancers , 2018, Science.

[144]  H. Clevers,et al.  Long-Term Adult Feline Liver Organoid Cultures for Disease Modeling of Hepatic Steatosis , 2017, Stem cell reports.

[145]  C. Coyne,et al.  Stem Cell-Derived Models of Viral Infections in the Gastrointestinal Tract , 2018, Viruses.

[146]  P. Garcez,et al.  Zika virus impairs growth in human neurospheres and brain organoids , 2016, Science.

[147]  S. Bartfeld Modeling infectious diseases and host-microbe interactions in gastrointestinal organoids. , 2016, Developmental biology.

[148]  C. Chen,et al.  Disease Modeling and Gene Therapy of Copper Storage Disease in Canine Hepatic Organoids , 2015, Stem cell reports.

[149]  B. Giepmans,et al.  Long-Term In Vitro Expansion of Salivary Gland Stem Cells Driven by Wnt Signals , 2015, Stem cell reports.

[150]  M. Eiraku,et al.  Functional anterior pituitary generated in self-organizing culture of human embryonic stem cells , 2016, Nature Communications.

[151]  D. Geelen,et al.  In Vitro Propagation , 2013 .

[152]  A. Oudenaarden,et al.  Long‐term expanding human airway organoids for disease modeling , 2019, The EMBO journal.

[153]  Kimberly A. Homan,et al.  Flow-enhanced vascularization and maturation of kidney organoids in vitro , 2018, Nature Methods.

[154]  Hans Clevers,et al.  Organoid Cultures Derived from Patients with Advanced Prostate Cancer , 2014, Cell.

[155]  Rebecca M Willett,et al.  Engineering induction of singular neural rosette emergence within hPSC-derived tissues , 2018, eLife.

[156]  Elizabeth E. Hoskins,et al.  Directed differentiation of human pluripotent stem cells into intestinal tissue in vitro , 2010, Nature.

[157]  E. Stanley,et al.  Directing human embryonic stem cell differentiation towards a renal lineage generates a self-organizing kidney , 2013, Nature Cell Biology.

[158]  Junjie Zhu,et al.  Inflammatory Cytokine TNFα Promotes the Long-Term Expansion of Primary Hepatocytes in 3D Culture , 2018, Cell.

[159]  M. Witjes,et al.  Human Salivary Gland Stem Cells Functionally Restore Radiation Damaged Salivary Glands , 2016, Stem cells.

[160]  M. Takasato,et al.  Understanding kidney morphogenesis to guide renal tissue regeneration , 2016, Nature Reviews Nephrology.

[161]  Katja Schenke-Layland,et al.  Self-Organized Cerebral Organoids with Human-Specific Features Predict Effective Drugs to Combat Zika Virus Infection. , 2017, Cell reports.

[162]  Teruo Fujii,et al.  Organ/body-on-a-chip based on microfluidic technology for drug discovery. , 2017, Drug metabolism and pharmacokinetics.

[163]  Marius Wernig,et al.  In vitro differentiation of transplantable neural precursors from human embryonic stem cells , 2001, Nature Biotechnology.

[164]  Mazen Asaad,et al.  A guide to using functional magnetic resonance imaging to study Alzheimer's disease in animal models , 2018, Disease Models & Mechanisms.

[165]  Hans Clevers,et al.  In vitro expansion of single Lgr5+ liver stem cells induced by Wnt-driven regeneration , 2013, Nature.

[166]  Eric D. Siggia,et al.  A method to recapitulate early embryonic spatial patterning in human embryonic stem cells , 2014, Nature Methods.

[167]  Hans Clevers,et al.  Disease Modeling in Stem Cell-Derived 3D Organoid Systems. , 2017, Trends in molecular medicine.

[168]  S. Nik-Zainal,et al.  Use of CRISPR-modified human stem cell organoids to study the origin of mutational signatures in cancer , 2017, Science.

[169]  J. C. Belmonte,et al.  Directed differentiation of human pluripotent cells to ureteric bud kidney progenitor-like cells , 2013, Nature Cell Biology.

[170]  E SMITH,et al.  Cystic organoid teratoma; report of a case. , 1946, Canadian Medical Association journal.

[171]  Yoshiki Sasai,et al.  Self-organized formation of polarized cortical tissues from ESCs and its active manipulation by extrinsic signals. , 2008, Cell stem cell.

[172]  Philippe Aubert,et al.  Engineered human pluripotent-stem-cell-derived intestinal tissues with a functional enteric nervous system , 2016, Nature Medicine.

[173]  G. Dontu,et al.  In vitro propagation and transcriptional profiling of human mammary stem/progenitor cells. , 2003, Genes & development.

[174]  J. Takagi,et al.  Human Pancreatic Tumor Organoids Reveal Loss of Stem Cell Niche Factor Dependence during Disease Progression. , 2018, Cell stem cell.

[175]  Z. Memish,et al.  Human intestinal tract serves as an alternative infection route for Middle East respiratory syndrome coronavirus , 2017, Science Advances.

[176]  Hans Clevers,et al.  In vitro expansion of human gastric epithelial stem cells and their responses to bacterial infection. , 2015, Gastroenterology.