Neurosphere based differentiation of human iPSC improves astrocyte differentiation

Shuling Zhou; Karolina Szczesna; Anna Ochalek; Julianna Kobolák; Eszter Varga; Csilla Nemes; Abinaya Chandrasekaran; Mikkel Rasmussen; Susanna Cirera Salicio; Poul Hyttel; András Dinnyés; Kristine Freude; Hasan X. Avci

doi:10.1155/2016/4937689

Neurosphere based differentiation of human iPSC improves astrocyte differentiation

Shuling Zhou, Karolina Szczesna, Anna Ochalek, Julianna Kobolák, Eszter Varga, Csilla Nemes, Abinaya Chandrasekaran, Mikkel Rasmussen, Susanna Cirera Salicio, Poul Hyttel, András Dinnyés, Kristine Freude, Hasan X. Avci

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Abstract

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.

Original language	English
Article number	4937689
Journal	Stem Cells International
Volume	2016
Number of pages	15
ISSN	1687-966X
DOIs	https://doi.org/10.1155/2016/4937689
Publication status	Published - 2016

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Neurosphere based differentiation of human iPSC improves astrocyte differentiationFinal published version, 2.61 MBLicence: CC BY

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title = "Neurosphere based differentiation of human iPSC improves astrocyte differentiation",

abstract = "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.",

author = "Shuling Zhou and Karolina Szczesna and Anna Ochalek and Julianna Kobol{\'a}k and Eszter Varga and Csilla Nemes and Abinaya Chandrasekaran and Mikkel Rasmussen and {Cirera Salicio}, Susanna and Poul Hyttel and Andr{\'a}s Dinny{\'e}s and Kristine Freude and Avci, {Hasan X.}",

year = "2016",

doi = "10.1155/2016/4937689",

language = "English",

volume = "2016",

journal = "Stem Cells International",

issn = "1687-966X",

publisher = "Hindawi Publishing Corporation",

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TY - JOUR

T1 - Neurosphere based differentiation of human iPSC improves astrocyte differentiation

AU - Zhou, Shuling

AU - Szczesna, Karolina

AU - Ochalek, Anna

AU - Kobolák, Julianna

AU - Varga, Eszter

AU - Nemes, Csilla

AU - Chandrasekaran, Abinaya

AU - Rasmussen, Mikkel

AU - Cirera Salicio, Susanna

AU - Hyttel, Poul

AU - Dinnyés, András

AU - Freude, Kristine

AU - Avci, Hasan X.

PY - 2016

Y1 - 2016

N2 - 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.

AB - 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.

U2 - 10.1155/2016/4937689

DO - 10.1155/2016/4937689

M3 - Journal article

C2 - 26798357

SN - 1687-966X

VL - 2016

JO - Stem Cells International

JF - Stem Cells International

M1 - 4937689

ER -

Neurosphere based differentiation of human iPSC improves astrocyte differentiation

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