Neurosphere Based Differentiation of Human iPSC Improves Astrocyte Differentiation

Neural progenitor cells (NPCs) derived from human induced pluripotent stem cells (iPSCs) are traditionally maintained and proliferated utilizing two-dimensional (2D) adherent monolayer culture systems. However, NPCs cultured using this system hardly reflect the intrinsic spatial development of brain...

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Main Authors: Shuling Zhou, Karolina Szczesna, Anna Ochalek, Julianna Kobolák, Eszter Varga, Csilla Nemes, Abinaya Chandrasekaran, Mikkel Rasmussen, Susanna Cirera, Poul Hyttel, András Dinnyés, Kristine K. Freude, Hasan X. Avci
Format: Article
Language:English
Published: Wiley 2016-01-01
Series:Stem Cells International
Online Access:http://dx.doi.org/10.1155/2016/4937689
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author Shuling Zhou
Karolina Szczesna
Anna Ochalek
Julianna Kobolák
Eszter Varga
Csilla Nemes
Abinaya Chandrasekaran
Mikkel Rasmussen
Susanna Cirera
Poul Hyttel
András Dinnyés
Kristine K. Freude
Hasan X. Avci
author_facet Shuling Zhou
Karolina Szczesna
Anna Ochalek
Julianna Kobolák
Eszter Varga
Csilla Nemes
Abinaya Chandrasekaran
Mikkel Rasmussen
Susanna Cirera
Poul Hyttel
András Dinnyés
Kristine K. Freude
Hasan X. Avci
author_sort Shuling Zhou
collection DOAJ
description Neural progenitor cells (NPCs) derived from human induced pluripotent stem cells (iPSCs) are traditionally maintained and proliferated utilizing two-dimensional (2D) adherent monolayer culture systems. However, NPCs cultured using this system hardly reflect the intrinsic spatial development of brain tissue. In this study, we determined that culturing iPSC-derived NPCs as three-dimensional (3D) floating neurospheres resulted in increased expression of the neural progenitor cell (NPC) markers, PAX6 and NESTIN. Expansion of NPCs in 3D culture methods also resulted in a more homogenous PAX6 expression when compared to 2D culture methods. Furthermore, the 3D propagation method for NPCs resulted in a significant higher expression of the astrocyte markers  GFAP and aquaporin 4 (AQP4) in the differentiated cells. Thus, our 3D propagation method could constitute a useful tool to promote NPC homogeneity and also to increase the differentiation potential of iPSC towards astrocytes.
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institution Kabale University
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spelling doaj-art-86b694ed24f8430ca16424a6ce94d5272025-08-20T03:37:53ZengWileyStem Cells International1687-966X1687-96782016-01-01201610.1155/2016/49376894937689Neurosphere Based Differentiation of Human iPSC Improves Astrocyte DifferentiationShuling Zhou0Karolina Szczesna1Anna Ochalek2Julianna Kobolák3Eszter Varga4Csilla Nemes5Abinaya Chandrasekaran6Mikkel Rasmussen7Susanna Cirera8Poul Hyttel9András Dinnyés10Kristine K. Freude11Hasan X. Avci12Department of Veterinary Clinical and Animal Science, Faculty of Health and Medical Sciences, University of Copenhagen, 1870 Frederiksberg, DenmarkBioTalentum Ltd., Gödöllo 2100, HungaryBioTalentum Ltd., Gödöllo 2100, HungaryBioTalentum Ltd., Gödöllo 2100, HungaryBioTalentum Ltd., Gödöllo 2100, HungaryBioTalentum Ltd., Gödöllo 2100, HungaryBioTalentum Ltd., Gödöllo 2100, HungaryBioneer A/S, 2970 Hørsholm, DenmarkDepartment of Veterinary Clinical and Animal Science, Faculty of Health and Medical Sciences, University of Copenhagen, 1870 Frederiksberg, DenmarkDepartment of Veterinary Clinical and Animal Science, Faculty of Health and Medical Sciences, University of Copenhagen, 1870 Frederiksberg, DenmarkBioTalentum Ltd., Gödöllo 2100, HungaryDepartment of Veterinary Clinical and Animal Science, Faculty of Health and Medical Sciences, University of Copenhagen, 1870 Frederiksberg, DenmarkBioTalentum Ltd., Gödöllo 2100, HungaryNeural progenitor cells (NPCs) derived from human induced pluripotent stem cells (iPSCs) are traditionally maintained and proliferated utilizing two-dimensional (2D) adherent monolayer culture systems. However, NPCs cultured using this system hardly reflect the intrinsic spatial development of brain tissue. In this study, we determined that culturing iPSC-derived NPCs as three-dimensional (3D) floating neurospheres resulted in increased expression of the neural progenitor cell (NPC) markers, PAX6 and NESTIN. Expansion of NPCs in 3D culture methods also resulted in a more homogenous PAX6 expression when compared to 2D culture methods. Furthermore, the 3D propagation method for NPCs resulted in a significant higher expression of the astrocyte markers  GFAP and aquaporin 4 (AQP4) in the differentiated cells. Thus, our 3D propagation method could constitute a useful tool to promote NPC homogeneity and also to increase the differentiation potential of iPSC towards astrocytes.http://dx.doi.org/10.1155/2016/4937689
spellingShingle Shuling Zhou
Karolina Szczesna
Anna Ochalek
Julianna Kobolák
Eszter Varga
Csilla Nemes
Abinaya Chandrasekaran
Mikkel Rasmussen
Susanna Cirera
Poul Hyttel
András Dinnyés
Kristine K. Freude
Hasan X. Avci
Neurosphere Based Differentiation of Human iPSC Improves Astrocyte Differentiation
Stem Cells International
title Neurosphere Based Differentiation of Human iPSC Improves Astrocyte Differentiation
title_full Neurosphere Based Differentiation of Human iPSC Improves Astrocyte Differentiation
title_fullStr Neurosphere Based Differentiation of Human iPSC Improves Astrocyte Differentiation
title_full_unstemmed Neurosphere Based Differentiation of Human iPSC Improves Astrocyte Differentiation
title_short Neurosphere Based Differentiation of Human iPSC Improves Astrocyte Differentiation
title_sort neurosphere based differentiation of human ipsc improves astrocyte differentiation
url http://dx.doi.org/10.1155/2016/4937689
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