Microencapsulation of dopamine neurons derived from human induced pluripotent stem cells

• Published in: Biochimica et biophysica acta Vol. 1850; no. 1; pp. 22 - 32
• Main Authors: Konagaya, Shuhei, Iwata, Hiroo
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.01.2015
• Subjects: agarose; cell aggregates; Cell Differentiation; Cells, Cultured; Cryopreservation; culture dishes; Cytological Techniques - methods; dopamine; Dopamine - secretion; Dopamine neuron; Dopaminergic Neurons - cytology; Dopaminergic Neurons - metabolism; Gene Expression; Humans; immune response; Induced pluripotent stem cell; induced pluripotent stem cells; Induced Pluripotent Stem Cells - cytology; Induced Pluripotent Stem Cells - metabolism; Microencapsulation; Microscopy, Fluorescence; Microspheres; neurons; Octamer Transcription Factor-3 - genetics; Parkinson disease; Pluripotent Stem Cells - cytology; Pluripotent Stem Cells - metabolism; Reverse Transcriptase Polymerase Chain Reaction; Sepharose; Technology, Pharmaceutical - methods; Time Factors; tyrosine; Tyrosine 3-Monooxygenase - genetics; Tyrosine 3-Monooxygenase - metabolism; Induced pluripotent stem cell; Dopamine neuron; Microencapsulation
• ISSN: 0304-4165; 0006-3002; 1872-8006
• DOI: 10.1016/j.bbagen.2014.09.025

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. 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 5days, cell aggregates were collected from the plates and microencapsulated in agarose microbeads. The microencapsulated aggregates were cultured for an additional 45days to induce maturation of dopamine neurons. Approximately 60% of all cells differentiated into tyrosine hydroxylase-positive neurons in agarose microbeads. The cells released dopamine for more than 40days. In addition, microbeads containing cells could be cryopreserved. hiPS cells were successfully differentiated into dopamine neurons in agarose microbeads. Agarose microencapsulation provides a good supporting environment for the preparation and storage of dopamine neurons. •Microencapsulation technique was used for the preparation of dopamine neurons.•HiPS-derived precursor cells were encapsulated in agarose microbeads.•Microencapsulated cells were successfully differentiated into dopamine neurons.•Microencapsulated cells could be cryopreserved.