Methods to extract and analyze fluid from human pluripotent stem cell-derived choroid plexus organoids

Leon H Chew; Eloi Mercier; Jason C Rogalski; Sara Pippard; Erin Knock

doi:10.3389/fnmol.2023.1243499

Methods to extract and analyze fluid from human pluripotent stem cell-derived choroid plexus organoids

Front Mol Neurosci. 2024 Jan 29:16:1243499. doi: 10.3389/fnmol.2023.1243499. eCollection 2023.

Authors

Leon H Chew¹, Eloi Mercier¹, Jason C Rogalski², Sara Pippard¹, Erin Knock¹

Affiliations

¹ STEMCELL Technologies, Vancouver, BC, Canada.
² Department of Biochemistry and Molecular Biology, University of British Columbia, Vancouver, BC, Canada.

Abstract

The choroid plexus (ChP) is a highly vascularized tissue lining the ventricular space of the brain. The ChP generates cerebrospinal fluid (CSF) and forms a protective barrier in the central nervous system (CNS). Recently, a three-dimensional human pluripotent stem cell (hPSC)-derived ChP organoid model has been developed. This model generates cystic structures that are filled with a fluid resembling CSF and are surrounded by an epithelial layer expressing ependymal choroid plexus-specific markers. Here we describe a method to generate these choroid plexus organoids using a commercial kit and methods to extract the CSF-like fluid for use in downstream analysis.

Keywords: cerebrospinal fluid; choroid plexus; mass spectrometry; organoid; organoid cell culture; pluripotent stem cell (PSC); proteomics.

Grants and funding

The authors declare that this study received funding from STEMCELL Technologies Inc. The funder was not involved in the study design, collection, analysis, interpretation of data, the writing of this article, or the decision to submit it for publication.