RRC ID 45331
Author Konagaya S, Iwata H.
Title Microencapsulation of dopamine neurons derived from human induced pluripotent stem cells.
Journal Biochim Biophys Acta
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.
CONCLUSION:hiPS cells were successfully differentiated into dopamine neurons in agarose microbeads.
GENERAL SIGNIFICANCE:Agarose microencapsulation provides a good supporting environment for the preparation and storage of dopamine neurons.
Volume 1850(1)
Pages 22-32
Published 2015-1-1
DOI 10.1016/j.bbagen.2014.09.025
PII S0304-4165(14)00327-4
PMID 25281770
MeSH Cell Differentiation* Cells, Cultured Cryopreservation Cytological Techniques / methods Dopamine / metabolism Dopaminergic Neurons / cytology Dopaminergic Neurons / metabolism* Gene Expression Humans Induced Pluripotent Stem Cells / cytology Induced Pluripotent Stem Cells / metabolism* Microscopy, Fluorescence Microspheres Octamer Transcription Factor-3 / genetics Pluripotent Stem Cells / cytology Pluripotent Stem Cells / metabolism Reverse Transcriptase Polymerase Chain Reaction Sepharose Technology, Pharmaceutical / methods Time Factors Tyrosine 3-Monooxygenase / genetics Tyrosine 3-Monooxygenase / metabolism*
IF 3.411
Times Cited 7
Human and Animal Cells 201B7(HPS0063) 253G1(HPS0002)