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Fluids and Barriers of the CNS

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Open Access 01-12-2016 | Research

Growth-factor reduced Matrigel source influences stem cell derived brain microvascular endothelial cell barrier properties

Authors: Ronak Patel, Abraham J. Alahmad

Published in: Fluids and Barriers of the CNS | Issue 1/2016

Abstract

Background

Patient-derived induced pluripotent stem cells (iPSCs) are an innovative source as an in vitro model for neurological diseases. Recent studies have demonstrated the differentiation of brain microvascular endothelial cells (BMECs) from various stem cell sources, including iPSC lines. However, the impact of the culturing conditions used to maintain such stem cell pluripotency on their ability to differentiate into BMECs remains undocumented. In this study, we investigated the effect of different sources of Matrigel and stem cell maintenance medium on BMEC differentiation efficiency.

Methods

The IMR90-c4 iPSC line was maintained on mTeSR1 or in essential-8 (E-8) medium on growth factor-reduced (GFR) Matrigel from three different manufacturers. Cells were differentiated into BMECs following published protocols. The phenotype of BMEC monolayers was assessed by immunocytochemistry. Barrier function was assessed by transendothelial electrical resistance (TEER) and permeability to sodium fluorescein, whereas the presence of drug efflux pumps was assessed by uptake assay using fluorescent substrates.

Results

Stem cell maintenance medium had little effect on the yield and barrier phenotype of IMR90-derived BMECs. The source of GFR-Matrigel used for the differentiation process significantly impacted the ability of IMR90-derived BMECs to form tight monolayers, as measured by TEER and fluorescein permeability. However, the Matrigel source had minimal effect on BMEC phenotype and drug efflux pump activity.

Conclusion

This study supports the ability to differentiate BMECs from iPSCs grown in mTeSR1 or E-8 medium and also suggests that the origin of GFR-Matrigel has a marked inpact on BMEC barrier properties.

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Title: Growth-factor reduced Matrigel source influences stem cell derived brain microvascular endothelial cell barrier properties
Authors: Ronak Patel
Abraham J. Alahmad
Publication date: 01-12-2016
Publisher: BioMed Central
Published in: Fluids and Barriers of the CNS / Issue 1/2016
Electronic ISSN: 2045-8118
DOI: https://doi.org/10.1186/s12987-016-0030-5

Keynote webinar | Spotlight on sleep in brain health

Springer Medicine

Growth-factor reduced Matrigel source influences stem cell derived brain microvascular endothelial cell barrier properties

Abstract

Background

Methods

Results

Conclusion

Keynote webinar | Spotlight on sleep in brain health

Springer Medicine

Abstract

Background

Methods

Results

Conclusion

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