Reproducible and efficient generation of functionally active neurons from human hiPSCs for preclinical disease modeling
The use of human induced pluripotent stem cell (hiPSC)-derived neuronal cultures to study the mechanisms of neurological disorders is often limited by low efficiency and high variability in differentiation of functional neurons. Here we compare the functional properties of neurons in cultures prepar...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2018-01-01
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| Series: | Stem Cell Research |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S1873506117302623 |
| _version_ | 1819163476253212672 |
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| author | Yunyao Xie Ryan J. Schutte Nathan N. Ng Kevin C. Ess Philip H. Schwartz Diane K. O'Dowd |
| author_facet | Yunyao Xie Ryan J. Schutte Nathan N. Ng Kevin C. Ess Philip H. Schwartz Diane K. O'Dowd |
| author_sort | Yunyao Xie |
| collection | DOAJ |
| description | The use of human induced pluripotent stem cell (hiPSC)-derived neuronal cultures to study the mechanisms of neurological disorders is often limited by low efficiency and high variability in differentiation of functional neurons. Here we compare the functional properties of neurons in cultures prepared with two hiPSC differentiation protocols, both plated on astroglial feeder layers. Using a protocol with an expandable intermediate stage, only a small percentage of cells with neuronal morphology were excitable by 21–23 days in culture. In contrast, a direct differentiation strategy of the same hiPSC line produced cultures in which the majority of neurons fired action potentials as early as 4–5 days. By 35–38 days over 80% of the neurons fired repetitively and many fired spontaneously. Spontaneous post-synaptic currents were observed in ~40% of the neurons at 4–5 days and in ~80% by 21–23 days. The majority (75%) received both glutamatergic and GABAergic spontaneous postsynaptic currents. The rate and degree of maturation of excitability and synaptic activity was similar between multiple independent platings from a single hiPSC line, and between two different control hiPSC lines. Cultures of rapidly functional neurons will facilitate identification of cellular mechanisms underlying genetically defined neurological disorders and development of novel therapeutics. Keywords: Induced pluripotent stem cells, hiPSC-derived neurons, Functional differentiation, Reproducibility, Disease modeling |
| first_indexed | 2024-12-22T17:44:44Z |
| format | Article |
| id | doaj.art-726e5342292e494fb341f65e0d078815 |
| institution | Directory Open Access Journal |
| issn | 1873-5061 |
| language | English |
| last_indexed | 2024-12-22T17:44:44Z |
| publishDate | 2018-01-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Stem Cell Research |
| spelling | doaj.art-726e5342292e494fb341f65e0d0788152022-12-21T18:18:19ZengElsevierStem Cell Research1873-50612018-01-01268494Reproducible and efficient generation of functionally active neurons from human hiPSCs for preclinical disease modelingYunyao Xie0Ryan J. Schutte1Nathan N. Ng2Kevin C. Ess3Philip H. Schwartz4Diane K. O'Dowd5Department of Developmental and Cell Biology, University of California, Irvine, Irvine, CA, United StatesDepartment of Developmental and Cell Biology, University of California, Irvine, Irvine, CA, United StatesDepartment of Developmental and Cell Biology, University of California, Irvine, Irvine, CA, United StatesDepartment of Pediatrics, Vanderbilt University Medical Center, Nashville, TN, United StatesChildren's Hospital of Orange County Research Institute, Orange, CA, United StatesDepartment of Developmental and Cell Biology, University of California, Irvine, Irvine, CA, United States; Corresponding author at: 4221 McGaugh Hall, University of California, Irvine, Irvine, CA 92697, United States.The use of human induced pluripotent stem cell (hiPSC)-derived neuronal cultures to study the mechanisms of neurological disorders is often limited by low efficiency and high variability in differentiation of functional neurons. Here we compare the functional properties of neurons in cultures prepared with two hiPSC differentiation protocols, both plated on astroglial feeder layers. Using a protocol with an expandable intermediate stage, only a small percentage of cells with neuronal morphology were excitable by 21–23 days in culture. In contrast, a direct differentiation strategy of the same hiPSC line produced cultures in which the majority of neurons fired action potentials as early as 4–5 days. By 35–38 days over 80% of the neurons fired repetitively and many fired spontaneously. Spontaneous post-synaptic currents were observed in ~40% of the neurons at 4–5 days and in ~80% by 21–23 days. The majority (75%) received both glutamatergic and GABAergic spontaneous postsynaptic currents. The rate and degree of maturation of excitability and synaptic activity was similar between multiple independent platings from a single hiPSC line, and between two different control hiPSC lines. Cultures of rapidly functional neurons will facilitate identification of cellular mechanisms underlying genetically defined neurological disorders and development of novel therapeutics. Keywords: Induced pluripotent stem cells, hiPSC-derived neurons, Functional differentiation, Reproducibility, Disease modelinghttp://www.sciencedirect.com/science/article/pii/S1873506117302623 |
| spellingShingle | Yunyao Xie Ryan J. Schutte Nathan N. Ng Kevin C. Ess Philip H. Schwartz Diane K. O'Dowd Reproducible and efficient generation of functionally active neurons from human hiPSCs for preclinical disease modeling Stem Cell Research |
| title | Reproducible and efficient generation of functionally active neurons from human hiPSCs for preclinical disease modeling |
| title_full | Reproducible and efficient generation of functionally active neurons from human hiPSCs for preclinical disease modeling |
| title_fullStr | Reproducible and efficient generation of functionally active neurons from human hiPSCs for preclinical disease modeling |
| title_full_unstemmed | Reproducible and efficient generation of functionally active neurons from human hiPSCs for preclinical disease modeling |
| title_short | Reproducible and efficient generation of functionally active neurons from human hiPSCs for preclinical disease modeling |
| title_sort | reproducible and efficient generation of functionally active neurons from human hipscs for preclinical disease modeling |
| url | http://www.sciencedirect.com/science/article/pii/S1873506117302623 |
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