Comparison of 2D and 3D neural induction methods for the generation of neural progenitor cells from human induced pluripotent stem cells
Neural progenitor cells (NPCs) from human induced pluripotent stem cells (hiPSCs) are frequently induced using 3D culture methodologies however, it is unknown whether spheroid-based (3D) neural induction is actually superior to monolayer (2D) neural induction. Our aim was to compare the efficiency o...
| Main Authors: | , , , , , , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2017-12-01
|
| Series: | Stem Cell Research |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S1873506117302131 |
| _version_ | 1818364734313857024 |
|---|---|
| author | Abinaya Chandrasekaran Hasan X. Avci Anna Ochalek Lone N. Rösingh Kinga Molnár Lajos László Tamás Bellák Annamária Téglási Krisztina Pesti Arpad Mike Phetcharat Phanthong Orsolya Bíró Vanessa Hall Narisorn Kitiyanant Karl-Heinz Krause Julianna Kobolák András Dinnyés |
| author_facet | Abinaya Chandrasekaran Hasan X. Avci Anna Ochalek Lone N. Rösingh Kinga Molnár Lajos László Tamás Bellák Annamária Téglási Krisztina Pesti Arpad Mike Phetcharat Phanthong Orsolya Bíró Vanessa Hall Narisorn Kitiyanant Karl-Heinz Krause Julianna Kobolák András Dinnyés |
| author_sort | Abinaya Chandrasekaran |
| collection | DOAJ |
| description | Neural progenitor cells (NPCs) from human induced pluripotent stem cells (hiPSCs) are frequently induced using 3D culture methodologies however, it is unknown whether spheroid-based (3D) neural induction is actually superior to monolayer (2D) neural induction. Our aim was to compare the efficiency of 2D induction with 3D induction method in their ability to generate NPCs, and subsequently neurons and astrocytes. Neural differentiation was analysed at the protein level qualitatively by immunocytochemistry and quantitatively by flow cytometry for NPC (SOX1, PAX6, NESTIN), neuronal (MAP2, TUBB3), cortical layer (TBR1, CUX1) and glial markers (SOX9, GFAP, AQP4). Electron microscopy demonstrated that both methods resulted in morphologically similar neural rosettes. However, quantification of NPCs derived from 3D neural induction exhibited an increase in the number of PAX6/NESTIN double positive cells and the derived neurons exhibited longer neurites. In contrast, 2D neural induction resulted in more SOX1 positive cells. While 2D monolayer induction resulted in slightly less mature neurons, at an early stage of differentiation, the patch clamp analysis failed to reveal any significant differences between the electrophysiological properties between the two induction methods. In conclusion, 3D neural induction increases the yield of PAX6+/NESTIN+ cells and gives rise to neurons with longer neurites, which might be an advantage for the production of forebrain cortical neurons, highlighting the potential of 3D neural induction, independent of iPSCs' genetic background. |
| first_indexed | 2024-12-13T22:09:04Z |
| format | Article |
| id | doaj.art-cab47fd19054428793390b890e26170a |
| institution | Directory Open Access Journal |
| issn | 1873-5061 1876-7753 |
| language | English |
| last_indexed | 2024-12-13T22:09:04Z |
| publishDate | 2017-12-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Stem Cell Research |
| spelling | doaj.art-cab47fd19054428793390b890e26170a2022-12-21T23:29:45ZengElsevierStem Cell Research1873-50611876-77532017-12-0125C13915110.1016/j.scr.2017.10.010Comparison of 2D and 3D neural induction methods for the generation of neural progenitor cells from human induced pluripotent stem cellsAbinaya Chandrasekaran0Hasan X. Avci1Anna Ochalek2Lone N. Rösingh3Kinga Molnár4Lajos László5Tamás Bellák6Annamária Téglási7Krisztina Pesti8Arpad Mike9Phetcharat Phanthong10Orsolya Bíró11Vanessa Hall12Narisorn Kitiyanant13Karl-Heinz Krause14Julianna Kobolák15András Dinnyés16BioTalentum Ltd, Gödöllő, HungaryBioTalentum Ltd, Gödöllő, HungaryBioTalentum Ltd, Gödöllő, HungaryDepartment of Pathology and Immunology, University of Geneva Medical School, Geneva, SwitzerlandDepartment of Anatomy, Cell and Developmental Biology, Eötvös Loránd University, Budapest, HungaryDepartment of Anatomy, Cell and Developmental Biology, Eötvös Loránd University, Budapest, HungaryBioTalentum Ltd, Gödöllő, HungaryBioTalentum Ltd, Gödöllő, HungaryOpto-Neuropharmacology Group, MTA-ELTE NAP B, Budapest, HungaryOpto-Neuropharmacology Group, MTA-ELTE NAP B, Budapest, HungaryBioTalentum Ltd, Gödöllő, HungaryFirst Department of Obstetrics and Gynaecology, Semmelweis University, Budapest, HungaryDepartment of Veterinary and Animal Science, University of Copenhagen, DenmarkStem Cell Research Group, Institute of Molecular Biosciences, Mahidol University, Nakhon Pathom Bangkok, ThailandDepartment of Pathology and Immunology, University of Geneva Medical School, Geneva, SwitzerlandBioTalentum Ltd, Gödöllő, HungaryBioTalentum Ltd, Gödöllő, HungaryNeural progenitor cells (NPCs) from human induced pluripotent stem cells (hiPSCs) are frequently induced using 3D culture methodologies however, it is unknown whether spheroid-based (3D) neural induction is actually superior to monolayer (2D) neural induction. Our aim was to compare the efficiency of 2D induction with 3D induction method in their ability to generate NPCs, and subsequently neurons and astrocytes. Neural differentiation was analysed at the protein level qualitatively by immunocytochemistry and quantitatively by flow cytometry for NPC (SOX1, PAX6, NESTIN), neuronal (MAP2, TUBB3), cortical layer (TBR1, CUX1) and glial markers (SOX9, GFAP, AQP4). Electron microscopy demonstrated that both methods resulted in morphologically similar neural rosettes. However, quantification of NPCs derived from 3D neural induction exhibited an increase in the number of PAX6/NESTIN double positive cells and the derived neurons exhibited longer neurites. In contrast, 2D neural induction resulted in more SOX1 positive cells. While 2D monolayer induction resulted in slightly less mature neurons, at an early stage of differentiation, the patch clamp analysis failed to reveal any significant differences between the electrophysiological properties between the two induction methods. In conclusion, 3D neural induction increases the yield of PAX6+/NESTIN+ cells and gives rise to neurons with longer neurites, which might be an advantage for the production of forebrain cortical neurons, highlighting the potential of 3D neural induction, independent of iPSCs' genetic background.http://www.sciencedirect.com/science/article/pii/S1873506117302131Neural inductionNeural progenitor cellshiPSCElectron microscopyPatch clamp2D-3D neural induction |
| spellingShingle | Abinaya Chandrasekaran Hasan X. Avci Anna Ochalek Lone N. Rösingh Kinga Molnár Lajos László Tamás Bellák Annamária Téglási Krisztina Pesti Arpad Mike Phetcharat Phanthong Orsolya Bíró Vanessa Hall Narisorn Kitiyanant Karl-Heinz Krause Julianna Kobolák András Dinnyés Comparison of 2D and 3D neural induction methods for the generation of neural progenitor cells from human induced pluripotent stem cells Stem Cell Research Neural induction Neural progenitor cells hiPSC Electron microscopy Patch clamp 2D-3D neural induction |
| title | Comparison of 2D and 3D neural induction methods for the generation of neural progenitor cells from human induced pluripotent stem cells |
| title_full | Comparison of 2D and 3D neural induction methods for the generation of neural progenitor cells from human induced pluripotent stem cells |
| title_fullStr | Comparison of 2D and 3D neural induction methods for the generation of neural progenitor cells from human induced pluripotent stem cells |
| title_full_unstemmed | Comparison of 2D and 3D neural induction methods for the generation of neural progenitor cells from human induced pluripotent stem cells |
| title_short | Comparison of 2D and 3D neural induction methods for the generation of neural progenitor cells from human induced pluripotent stem cells |
| title_sort | comparison of 2d and 3d neural induction methods for the generation of neural progenitor cells from human induced pluripotent stem cells |
| topic | Neural induction Neural progenitor cells hiPSC Electron microscopy Patch clamp 2D-3D neural induction |
| url | http://www.sciencedirect.com/science/article/pii/S1873506117302131 |
| work_keys_str_mv | AT abinayachandrasekaran comparisonof2dand3dneuralinductionmethodsforthegenerationofneuralprogenitorcellsfromhumaninducedpluripotentstemcells AT hasanxavci comparisonof2dand3dneuralinductionmethodsforthegenerationofneuralprogenitorcellsfromhumaninducedpluripotentstemcells AT annaochalek comparisonof2dand3dneuralinductionmethodsforthegenerationofneuralprogenitorcellsfromhumaninducedpluripotentstemcells AT lonenrosingh comparisonof2dand3dneuralinductionmethodsforthegenerationofneuralprogenitorcellsfromhumaninducedpluripotentstemcells AT kingamolnar comparisonof2dand3dneuralinductionmethodsforthegenerationofneuralprogenitorcellsfromhumaninducedpluripotentstemcells AT lajoslaszlo comparisonof2dand3dneuralinductionmethodsforthegenerationofneuralprogenitorcellsfromhumaninducedpluripotentstemcells AT tamasbellak comparisonof2dand3dneuralinductionmethodsforthegenerationofneuralprogenitorcellsfromhumaninducedpluripotentstemcells AT annamariateglasi comparisonof2dand3dneuralinductionmethodsforthegenerationofneuralprogenitorcellsfromhumaninducedpluripotentstemcells AT krisztinapesti comparisonof2dand3dneuralinductionmethodsforthegenerationofneuralprogenitorcellsfromhumaninducedpluripotentstemcells AT arpadmike comparisonof2dand3dneuralinductionmethodsforthegenerationofneuralprogenitorcellsfromhumaninducedpluripotentstemcells AT phetcharatphanthong comparisonof2dand3dneuralinductionmethodsforthegenerationofneuralprogenitorcellsfromhumaninducedpluripotentstemcells AT orsolyabiro comparisonof2dand3dneuralinductionmethodsforthegenerationofneuralprogenitorcellsfromhumaninducedpluripotentstemcells AT vanessahall comparisonof2dand3dneuralinductionmethodsforthegenerationofneuralprogenitorcellsfromhumaninducedpluripotentstemcells AT narisornkitiyanant comparisonof2dand3dneuralinductionmethodsforthegenerationofneuralprogenitorcellsfromhumaninducedpluripotentstemcells AT karlheinzkrause comparisonof2dand3dneuralinductionmethodsforthegenerationofneuralprogenitorcellsfromhumaninducedpluripotentstemcells AT juliannakobolak comparisonof2dand3dneuralinductionmethodsforthegenerationofneuralprogenitorcellsfromhumaninducedpluripotentstemcells AT andrasdinnyes comparisonof2dand3dneuralinductionmethodsforthegenerationofneuralprogenitorcellsfromhumaninducedpluripotentstemcells |