title: Microencapsulation of dopamine neurons derived from human induced pluripotent stem cells.
abstract: BACKGROUND
  Dopamine neurons derived from induced pluripotent stem cells have been widely studied for the treatment of Parkinson's disease. However, various difficulties remain to be overcome, such as tumor formation, fragility of dopamine neurons, difficulty in handling large numbers of dopamine neurons, and immune reactions. In this study, human induced pluripotent stem cell-derived precursors of dopamine neurons were encapsulated in agarose microbeads. Dopamine neurons in microbeads could be handled without specific protocols, because the microbeads protected the fragile dopamine neurons from mechanical stress.
  
  
  METHODS
  hiPS cells were seeded on a Matrigel-coated dish and cultured to induce differentiation into a dopamine neuronal linage. On day 18 of culture, cells were collected from the culture dishes and seeded into U-bottom 96-well plates to induce cell aggregate formation. After 5 days, cell aggregates were collected from the plates and microencapsulated in agarose microbeads. The microencapsulated aggregates were cultured for an additional 45 days to induce maturation of dopamine neurons.
  
  
  RESULTS
  Approximately 60% of all cells differentiated into tyrosine hydroxylase-positive neurons in agarose microbeads. The cells released dopamine for more than 40 days. In addition, microbeads containing cells could be cryopreserved.
  
  
  CONCLUSIONS
  hiPS cells were successfully differentiated into dopamine neurons in agarose microbeads.
  
  
  CONCLUSIONS
  Agarose microencapsulation provides a good supporting environment for the preparation and storage of dopamine neurons.
authors.affiliation: Institute for Frontier Medical Sciences, Kyoto University, 53 Kawahara-cho, Shogoin, Sakyo-ku, Kyoto 606-8507, Japan.
authors.forename: Shuhei
authors.initials: S
authors.lastname: Konagaya
authors.affiliation: Institute for Frontier Medical Sciences, Kyoto University, 53 Kawahara-cho, Shogoin, Sakyo-ku, Kyoto 606-8507, Japan. Electronic address: iwata@frontier.kuoyo-u.ac.jp.
authors.forename: Hiroo
authors.initials: H
authors.lastname: Iwata
mesh_terms: D002454:Cell Differentiation; D002478:Cells, Cultured; D015925:Cryopreservation; D003584:Cytological Techniques / Q000379:methods; D004298:Dopamine / Q000378:metabolism; D059290:Dopaminergic Neurons / Q000166:cytology / Q000378:metabolism*; D015870:Gene Expression; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D008863:Microspheres; D050814:Octamer Transcription Factor-3 / Q000235:genetics; D039904:Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012685:Sepharose; D013678:Technology, Pharmaceutical / Q000379:methods; D013997:Time Factors; D014446:Tyrosine 3-Monooxygenase / Q000235:genetics / Q000378:metabolism*
chemical_list: D050814:Octamer Transcription Factor-3; C494173:POU5F1 protein, human; D012685:Sepharose; D014446:Tyrosine 3-Monooxygenase; D004298:Dopamine
keywords: Dopamine neuron; Induced pluripotent stem cell; Microencapsulation