Comparison of 2D and 3D neural induction methods for the generation of neural progenitor cells from human induced pluripotent stem cells
| Autor: | Lajos László, Karl-Heinz Krause, Krisztina Pesti, Abinaya Chandrasekaran, Arpad Mike, Phetcharat Phanthong, Julianna Kobolák, Tamás Bellák, Orsolya Biró, Lone N. Rösingh, Vanessa Jane Hall, Narisorn Kitiyanant, Hasan X. Avci, Andras Dinnyes, Kinga Molnár, Annamária Téglási, Anna Ochalek |
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| Jazyk: | angličtina |
| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
PAX6 Transcription Factor Neurite Induced Pluripotent Stem Cells Cell Culture Techniques 2D-3D neural induction Cell Culture Techniques/methods Biology ddc:616.07 Neural Stem Cells/cytology/metabolism hiPSC Cell Line PAX6 Transcription Factor/genetics/metabolism Nestin 03 medical and health sciences SOX1 Neural Stem Cells Neural induction Neurosphere Electron microscopy Humans Neural progenitor cells Induced pluripotent stem cell lcsh:QH301-705.5 Nestin/genetics/metabolism SOXB1 Transcription Factors Induced Pluripotent Stem Cells/cytology/metabolism Cell Differentiation Cell Biology General Medicine Neural stem cell Cell biology Neuroepithelial cell 030104 developmental biology lcsh:Biology (General) nervous system SOXB1 Transcription Factors/genetics/metabolism Immunology Patch clamp Neural development Developmental Biology |
| Zdroj: | Stem Cell Research, Vol. 25 (2017) pp. 139-151 Stem Cell Research Chandrasekaran, A, Avci, H, Ochalek, A, Rosingh, L, Molnar, K, Laszlo, L, Bellak, T, Teglasi, A, Pesti, K, Mike, A, Phanthong, P, Biro, O, Hall, V J, Kitiyanant, N, Krause, K-H, Kobolak, J & Dinnyés, A 2017, ' Comparison of 2D and 3D neural induction methods for the generation of neural progenitor cells from human induced pluripotent stem cells ', Stem Cell Research, vol. 25, pp. 139-151 . https://doi.org/10.1016/j.scr.2017.10.010 Stem Cell Research, Vol 25, Iss C, Pp 139-151 (2017) |
| ISSN: | 1873-5061 |
| DOI: | 10.1016/j.scr.2017.10.010 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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