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Enhanced xeno-free differentiation of hiPSC-derived astroglia applied in a blood-brain barrier model

Journal article
Authors Louise Delsing
T. Kallur
Henrik Zetterberg
R. Hicks
J. Synnergren
Published in Fluids and Barriers of the Cns
Volume 16
Issue 1
ISSN 2045-8118
Publication year 2019
Published at Institute of Neuroscience and Physiology, Department of Psychiatry and Neurochemistry
Language en
Links dx.doi.org/10.1186/s12987-019-0147-...
Keywords Astroglia, hiPSC, In vitro models, Differentiation, Laminin-521, Blood-brain barrier, pluripotent stem-cells, endothelial-cells, directed differentiation, laminin, angiopoietin-1, expression, neurons, establishment, derivation, astrocytes, Neurosciences & Neurology, houck mp, 1990, journal of neurochemistry, v54, p1798
Subject categories Neurosciences

Abstract

Background Human induced pluripotent stem cells (hiPSC) hold great promise for use in cell therapy applications and for improved in vitro models of human disease. So far, most hiPSC differentiation protocols to astroglia use undefined, animal-containing culture matrices. Laminins, which play an essential role in the regulation of cell behavior, offer a source of defined, animal-free culture matrix. Methods In order to understand how laminins affect astroglia differentiation, recombinant human laminin-521 (LN521), was compared to a murine Engelbreth-Holm-Swarm sarcoma derived laminin (L2020). Astroglia expression of protein and mRNA together with glutamate uptake and protein secretion function, were evaluated. Finally, these astroglia were evaluated in a coculture model of the blood-brain barrier (BBB). Results Astroglia of good quality were generated from hiPSC on both LN521 and L2020. However, astroglia differentiated on human LN521 showed higher expression of several astroglia specific mRNAs and proteins such as GFAP, S100B, Angiopoietin-1, and EAAT1, compared to astroglia differentiated on murine L2020. In addition, glutamate uptake and ability to induce expression of junction proteins in endothelial cells were affected by the culture matrix for differentiation. Conclusion Our results suggest that astroglia differentiated on LN521 display an improved phenotype and are suitable for coculture in a hiPSC-derived BBB model. This provides a starting point for a more defined and robust derivation of astroglia for use in BBB coculture models.

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