TY - JOUR AU - Chandrasekaran, A AU - Avci, HX AU - Ochalek, A AU - Rosingh, LN AU - Molnár, Kinga AU - László, Lajos AU - Bellák, Tamás AU - Teglasi, A AU - Pesti, Krisztina AU - Mike, Árpád AU - Phanthong, P AU - Biró, Orsolya AU - Hall, V AU - Kitiyanant, N AU - Krause, KH AU - Kobolák, Julianna AU - Dinnyés, András TI - Comparison of 2D and 3D neural induction methods for the generation of neural progenitor cells from human induced pluripotent stem cells JF - STEM CELL RESEARCH J2 - STEM CELL RES VL - 25 PY - 2017 SP - 139 EP - 151 PG - 13 SN - 1873-5061 DO - 10.1016/j.scr.2017.10.010 UR - https://m2.mtmt.hu/api/publication/3312483 ID - 3312483 N1 - BioTalentum Ltd, Gödöllő, Hungary Department of Anatomy, Embryology and Histology, Faculty of Medicine, University of Szeged, Szeged, Hungary Molecular Animal Biotechnology Lab, Szent István University, Gödöllő, Hungary Department of Pathology and Immunology, University of Geneva Medical School, Geneva, Switzerland Department of Anatomy, Cell and Developmental Biology, Eötvös Loránd University, Budapest, Hungary Opto-Neuropharmacology Group, MTA-ELTE NAP B, Budapest, Hungary János Szentágothai Doctoral School of Neurosciences, Semmelweis University, Budapest, Hungary Stem Cell Research Group, Institute of Molecular Biosciences, Mahidol University, Nakhon Pathom Bangkok, Thailand First Department of Obstetrics and Gynaecology, Semmelweis University, Budapest, Hungary Department of Veterinary and Animal Science, University of Copenhagen, Denmark Cited By :88 Export Date: 9 April 2024 Correspondence Address: Dinnyés, A.; BioTalentum LtdHungary; email: Manuscript.Dinnyes@biotalentum.hu AB - 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. LA - English DB - MTMT ER - TY - JOUR AU - Xu, T AU - Molnár, Péter AU - Gregory, C AU - Das, M AU - Boland, T AU - Hickman, JJ TI - Electrophysiological characterization of embryonic hippocampal neurons cultured in a 3D collagen hydrogel JF - BIOMATERIALS J2 - BIOMATERIALS VL - 30 PY - 2009 IS - 26 SP - 4377 EP - 4383 PG - 7 SN - 0142-9612 DO - 10.1016/j.biomaterials.2009.04.047 UR - https://m2.mtmt.hu/api/publication/1789506 ID - 1789506 N1 - Megjegyzés-21884966 Z9: 5 WC: Engineering, Biomedical; Materials Science, Biomaterials Megjegyzés-21885170 Z9: 5 WC: Engineering, Biomedical; Materials Science, Biomaterials Megjegyzés-21885186 Z9: 5 WC: Engineering, Biomedical; Materials Science, Biomaterials Megjegyzés-21885258 Z9: 5 WC: Engineering, Biomedical; Materials Science, Biomaterials AB - Rat embryonic hippocampal neurons were cultured in (1) 3D collagen hydrogels as 'entrapped' evenly distributed cells, (2) at the interface of two collagen layers (sandwich model), and (3) on the surface of collagen coated coverslips (2D model). In the 'entrapment' model the neuronal processes grew out of the plane of the cell body and extended into the collagen matrix, in contrast to the sandwich model where the cells and their processes rarely left the plane in which they were seeded. Hippocampal neurons 'entrapped' in the 3D collagen gel grew the same number, but shorter, processes and exhibited improved survival compared to neurons cultured in the 2D model. There was no difference in the electrophysiological properties of the neurons cultured in the 3D compared to the 2D model except in the resting membrane potential and in the duration of the after-hyperpolarization. Spontaneous postsynaptic currents were recorded in 14- and 21-day-old 3D cultures evidencing functional synapse formation. Our results indicate that the physiological characteristics of 3D neuronal cultures are similar to traditional 2D cultures. However, functional 3D networks of hippocampal neurons will be necessary for multi-level circuit formation, which could be essential for understanding the basis of physiological learning and memory. (c) 2009 Elsevier Ltd. All rights reserved. LA - English DB - MTMT ER - TY - JOUR AU - Xu, T AU - Gregory, CA AU - Molnár, Péter AU - Cui, X AU - Jalota, S AU - Bhaduri, SB AU - Boland, T TI - Viability and electrophysiology of neural cell structures generated by the inkjet printing method JF - BIOMATERIALS J2 - BIOMATERIALS VL - 27 PY - 2006 IS - 19 SP - 3580 EP - 3588 PG - 9 SN - 0142-9612 DO - 10.1016/j.biomaterials.2006.01.048 UR - https://m2.mtmt.hu/api/publication/1789520 ID - 1789520 N1 - Z9: 76 WC: Engineering, Biomedical; Materials Science, Biomaterials AB - Complex cellular patterns and structures were created by automated and direct inkjet printing of primary embryonic hippocampal and cortical neurons. Immunostaining analysis and whole-cell patch-clamp recordings showed that embryonic hippocampal and cortical neurons maintained basic cellular properties and functions, including normal, healthy neuronal phenotypes and electrophysiological characteristics, after being printed through thermal inkjet nozzles. In addition, in this study a new method was developed to create 3D cellular structures: sheets of neural cells were layered on each other (layer-by-layer process) by alternate inkjet printing of NT2 cells and fibrin gels. These results and findings, taken together, show that inkjet printing is rapidly evolving into a digital fabrication method to build functional neural structures that may eventually find applications in neural tissue engineering. (c) 2006 Published by Elsevier Ltd. LA - English DB - MTMT ER -