title: Rapid Induction of Cerebral Organoids From Human Induced Pluripotent Stem Cells Using a Chemically Defined Hydrogel and Defined Cell Culture Medium.
abstract: Tissue organoids are a promising technology that may accelerate development of the societal and NIH mandate for precision medicine. Here we describe a robust and simple method for generating cerebral organoids (cOrgs) from human pluripotent stem cells by using a chemically defined hydrogel material and chemically defined culture medium. By using no additional neural induction components, cOrgs appeared on the hydrogel surface within 10-14 days, and under static culture conditions, they attained sizes up to 3 mm in greatest dimension by day 28. Histologically, the organoids showed neural rosette and neural tube-like structures and evidence of early corticogenesis. Immunostaining and quantitative reverse-transcription polymerase chain reaction demonstrated protein and gene expression representative of forebrain, midbrain, and hindbrain development. Physiologic studies showed responses to glutamate and depolarization in many cells, consistent with neural behavior. The method of cerebral organoid generation described here facilitates access to this technology, enables scalable applications, and provides a potential pathway to translational applications where defined components are desirable.
  
  
  CONCLUSIONS
  Tissue organoids are a promising technology with many potential applications, such as pharmaceutical screens and development of in vitro disease models, particularly for human polygenic conditions where animal models are insufficient. This work describes a robust and simple method for generating cerebral organoids from human induced pluripotent stem cells by using a chemically defined hydrogel material and chemically defined culture medium. This method, by virtue of its simplicity and use of defined materials, greatly facilitates access to cerebral organoid technology, enables scalable applications, and provides a potential pathway to translational applications where defined components are desirable.
authors.affiliation: Stem Cell Institute, University of Minnesota, Minneapolis, Minnesota, USA Bioactive Regenerative Therapeutics, Inc., Two Harbors, Minnesota, USA Department of Veterinary Population Medicine, University of Minnesota, St. Paul, Minnesota, USA.
authors.forename: Beth A
authors.initials: BA
authors.lastname: Lindborg
authors.affiliation: Bioactive Regenerative Therapeutics, Inc., Two Harbors, Minnesota, USA.
authors.forename: John H
authors.initials: JH
authors.lastname: Brekke
authors.affiliation: Stem Cell Institute, University of Minnesota, Minneapolis, Minnesota, USA Department of Veterinary Population Medicine, University of Minnesota, St. Paul, Minnesota, USA.
authors.forename: Amanda L
authors.initials: AL
authors.lastname: Vegoe
authors.affiliation: Stem Cell Institute, University of Minnesota, Minneapolis, Minnesota, USA Department of Veterinary Population Medicine, University of Minnesota, St. Paul, Minnesota, USA.
authors.forename: Connor B
authors.initials: CB
authors.lastname: Ulrich
authors.affiliation: Stem Cell Institute, University of Minnesota, Minneapolis, Minnesota, USA Department of Veterinary Population Medicine, University of Minnesota, St. Paul, Minnesota, USA.
authors.forename: Kerri T
authors.initials: KT
authors.lastname: Haider
authors.affiliation: Stem Cell Institute, University of Minnesota, Minneapolis, Minnesota, USA Department of Veterinary Population Medicine, University of Minnesota, St. Paul, Minnesota, USA Department of Neurosurgery, University of Minnesota, Minneapolis, Minnesota, USA.
authors.forename: Sandhya
authors.initials: S
authors.lastname: Subramaniam
authors.affiliation: Stem Cell Institute, University of Minnesota, Minneapolis, Minnesota, USA Department of Veterinary Population Medicine, University of Minnesota, St. Paul, Minnesota, USA.
authors.forename: Scott L
authors.initials: SL
authors.lastname: Venhuizen
authors.affiliation: Stem Cell Institute, University of Minnesota, Minneapolis, Minnesota, USA Department of Pediatrics, University of Minnesota, Minneapolis, Minnesota, USA.
authors.forename: Cindy R
authors.initials: CR
authors.lastname: Eide
authors.affiliation: Department of Pediatrics, University of Minnesota, Minneapolis, Minnesota, USA.
authors.forename: Paul J
authors.initials: PJ
authors.lastname: Orchard
authors.affiliation: Stem Cell Institute, University of Minnesota, Minneapolis, Minnesota, USA Department of Pediatrics, University of Minnesota, Minneapolis, Minnesota, USA.
authors.forename: Weili
authors.initials: W
authors.lastname: Chen
authors.affiliation: Biostatistical Design and Analysis Center, Clinical and Translational Science Institute, University of Minnesota, Minneapolis, Minnesota, USA.
authors.forename: Qi
authors.initials: Q
authors.lastname: Wang
authors.affiliation: Department of Chemical Engineering and Materials Science, University of Minnesota, Minneapolis, Minnesota, USA.
authors.forename: Francisco
authors.initials: F
authors.lastname: Pelaez
authors.affiliation: Department of Biomedical Engineering, University of Minnesota, Minneapolis, Minnesota, USA.
authors.forename: Carolyn M
authors.initials: CM
authors.lastname: Scott
authors.affiliation: Department of Chemical Engineering and Materials Science, University of Minnesota, Minneapolis, Minnesota, USA.
authors.forename: Efrosini
authors.initials: E
authors.lastname: Kokkoli
authors.affiliation: Stem Cell Institute, University of Minnesota, Minneapolis, Minnesota, USA Department of Integrative Biology and Physiology, University of Minnesota, Minneapolis, Minnesota, USA.
authors.forename: Susan A
authors.initials: SA
authors.lastname: Keirstead
authors.affiliation: Stem Cell Institute, University of Minnesota, Minneapolis, Minnesota, USA Department of Genetics, Cell Biology, and Development, University of Minnesota, Minneapolis, Minnesota, USA.
authors.forename: James R
authors.initials: JR
authors.lastname: Dutton
authors.affiliation: Stem Cell Institute, University of Minnesota, Minneapolis, Minnesota, USA Department of Pediatrics, University of Minnesota, Minneapolis, Minnesota, USA.
authors.forename: Jakub
authors.initials: J
authors.lastname: Tolar
authors.affiliation: Stem Cell Institute, University of Minnesota, Minneapolis, Minnesota, USA Department of Veterinary Population Medicine, University of Minnesota, St. Paul, Minnesota, USA obrie004@umn.edu.
authors.forename: Timothy D
authors.initials: TD
authors.lastname: O'Brien
mesh_terms: D001696:Biomechanical Phenomena; D001921:Brain / Q000166:cytology* / Q000378:metabolism; D002454:Cell Differentiation / Q000235:genetics; D002478:Cells, Cultured; D003470:Culture Media / Q000737:chemistry* / Q000494:pharmacology; D015870:Gene Expression; D006801:Humans; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate / Q000737:chemistry / Q000494:pharmacology*; D057026:Induced Pluripotent Stem Cells / Q000502:physiology*; D009474:Neurons / Q000166:cytology / Q000502:physiology; D009940:Organoids / Q000166:cytology / Q000502:physiology*; D046509:Tissue Culture Techniques / Q000379:methods*
chemical_list: D003470:Culture Media; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate
keywords: Adrenoleukodystrophy; Brain; In vitro techniques; Organoids; Stem cells