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 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 - Fuzik, J AU - Zeisel, A AU - Máté, Zoltán AU - Calvigioni, D AU - Yanagawa, Y AU - Szabó, Gábor AU - Linnarsson, S AU - Harkany, T TI - Integration of electrophysiological recordings with single-cell RNA-seq data identifies neuronal subtypes. JF - NATURE BIOTECHNOLOGY J2 - NAT BIOTECHNOL VL - 34 PY - 2016 IS - 2 SP - 175 EP - 183 PG - 9 SN - 1087-0156 DO - 10.1038/nbt.3443 UR - https://m2.mtmt.hu/api/publication/3208369 ID - 3208369 N1 - Department of Medical Biochemistry and Biophysics, Karolinska Institutet, Stockholm, Sweden Department of Molecular Neurosciences, Center for Brain Research, Medical University of Vienna, Vienna, Austria Institute of Experimental Medicine, Hungarian Academy of Sciences, Budapest, Hungary Department of Genetic and Behavioral Neuroscience, Gunma University, Graduate School of Medicine, Maebashi, Japan Cited By :246 Export Date: 9 June 2023 CODEN: NABIF Correspondence Address: Harkany, T.; Department of Medical Biochemistry and Biophysics, Sweden; email: tibor.harkany@meduniwien.ac.at Chemicals/CAS: cholecystokinin, 9011-97-6, 93443-27-7; gamma synuclein, 216864-09-4; synaptotagmin, 134193-26-3, 134193-27-4; RNA, Messenger Funding details: Seventh Framework Programme, FP7, 261063 Funding details: European Research Council, ERC Funding details: Human Frontier Science Program, HFSP Funding details: Vetenskapsrådet, VR Funding details: Novo Nordisk Fonden, NNF Funding text 1: We thank A. Juréus for DNA sequencing, and the CLICK Imaging Facility at Karolinska Institutet for making the Imaris software package available for neuronal reconstructions, T. Klausberger and E. Borók for discussions and assistance with Neurolucida reconstructions. This work was supported by the European Research Council (BRAINCELL 261063, to S.L.), the Swedish Research Council (to S.L. and T.H.); Human Frontier Science Program (to A.Z.), the European Commission 7th Framework Program (PAINCAGE, to T.H.), Hjärnfonden (to T.H.) and the NovoNordisk Foundation (to T.H.). AB - Traditionally, neuroscientists have defined the identity of neurons by the cells' location, morphology, connectivity and excitability. However, the direct relationship between these parameters and the molecular phenotypes has remained largely unexplored. Here, we present a method for obtaining full transcriptome data from single neocortical pyramidal cells and interneurons after whole-cell patch-clamp recordings in mouse brain slices. In our approach, termed Patch-seq, a patch-clamp stimulus protocol is followed by the aspiration of the entire somatic compartment into the recording pipette, reverse transcription of RNA including addition of unique molecular identifiers, cDNA amplification, Illumina library preparation and sequencing. We show that Patch-seq reveals a close link between electrophysiological characteristics, responses to acute chemical challenges and RNA expression of neurotransmitter receptors and channels. Moreover, it distinguishes neuronal subpopulations that correspond to both well-established and, to our knowledge, hitherto undescribed neuronal subtypes. Our findings demonstrate the ability of Patch-seq to precisely map neuronal subtypes and predict their network contributions in the brain. LA - English DB - MTMT ER - TY - JOUR AU - Mariani, J AU - Simonini, MV AU - Palejev, D AU - Tomasini, L AU - Coppola, G AU - Szekely, AM AU - Horváth, Tamás AU - Vaccarino, FM TI - Modeling human cortical development in vitro using induced pluripotent stem cells JF - PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA J2 - P NATL ACAD SCI USA VL - 109 PY - 2012 IS - 31 SP - 12770 EP - 12775 PG - 6 SN - 0027-8424 DO - 10.1073/pnas.1202944109 UR - https://m2.mtmt.hu/api/publication/3198991 ID - 3198991 LA - English DB - MTMT ER - TY - JOUR AU - Levine, AJ AU - Brivanlou, Ali H. TI - Proposal of a model of mammalian neural induction. JF - DEVELOPMENTAL BIOLOGY J2 - DEV BIOL VL - 308 PY - 2007 IS - 2 SP - 247 EP - 256 PG - 10 SN - 0012-1606 DO - 10.1016/j.ydbio.2007.05.036 UR - https://m2.mtmt.hu/api/publication/3250820 ID - 3250820 AB - How does the vertebrate embryo make a nervous system? This complex question has been at the center of developmental biology for many years. The earliest step in this process - the induction of neural tissue - is intimately linked to patterning of the entire early embryo, and the molecular and embryological of basis these processes are beginning to emerge. Here, we analyze classic and cutting-edge findings on neural induction in the mouse. We find that data from genetics, tissue explants, tissue grafting, and molecular marker expression support a coherent framework for mammalian neural induction. In this model, the gastrula organizer of the mouse embryo inhibits BMP signaling to allow neural tissue to form as a default fate-in the absence of instructive signals. The first neural tissue induced is anterior and subsequent neural tissue is posteriorized to form the midbrain, hindbrain, and spinal cord. The anterior visceral endoderm protects the pre-specified anterior neural fate from similar posteriorization, allowing formation of forebrain. This model is very similar to the default model of neural induction in the frog, thus bridging the evolutionary gap between amphibians and mammals. LA - English DB - MTMT ER - TY - JOUR AU - Barberi, T AU - Klivényi, Péter AU - Calingasan, NY AU - Lee, H AU - Kawamata, H AU - Loonam, K AU - Perrier, AL AU - Bruses, J AU - Rubio, ME AU - Topf, N AU - Tabar, V AU - Harrison, NL AU - Beal, MF AU - Moore, MA AU - Studer, L TI - Neural subtype specification of fertilization and nuclear transfer embryonic stem cells and application in parkinsonian mice JF - NATURE BIOTECHNOLOGY J2 - NAT BIOTECHNOL VL - 21 PY - 2003 IS - 10 SP - 1200 EP - 1207 PG - 8 SN - 1087-0156 DO - 10.1038/nbt870 UR - https://m2.mtmt.hu/api/publication/1410336 ID - 1410336 LA - English DB - MTMT ER - TY - JOUR AU - Munoz-Sanjuan, I AU - Brivanlou, Ali H. TI - Neural induction, the default model and embryonic stem cells. JF - NATURE REVIEWS NEUROSCIENCE J2 - NAT REV NEUROSCI VL - 3 PY - 2002 IS - 4 SP - 271 EP - 280 PG - 10 SN - 1471-003X DO - 10.1038/nrn786 UR - https://m2.mtmt.hu/api/publication/3250855 ID - 3250855 LA - English DB - MTMT ER -