Methods to Extract and Analyze Fluid from Human Pluripotent Stem Cell-derived Choroid Plexus Organoids

Overview

Journal Front Mol Neurosci

Specialty Molecular Biology

Date 2024 Feb 13

PMID 38348236

Authors

Leon H Chew

Eloi Mercier

Jason C Rogalski

Sara Pippard

Erin Knock

Affiliations

Soon will be listed here.

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.

Citing Articles

Brain organoid methodologies to explore mechanisms of disease in progressive multiple sclerosis.

Simoes-Abade M, Patterer M, Nicaise A, Pluchino S Front Cell Neurosci. 2025; 18:1488691.

PMID: 39744673 PMC: 11688374. DOI: 10.3389/fncel.2024.1488691.

References

Watanabe M, Kang Y, Davies L, Meghpara S, Lau K, Chung C . BMP4 sufficiency to induce choroid plexus epithelial fate from embryonic stem cell-derived neuroepithelial progenitors. J Neurosci. 2012; 32(45):15934-45. PMC: 3505486. DOI: 10.1523/JNEUROSCI.3227-12.2012. View

Chew L, Anonuevo A, Knock E . Generating Cerebral Organoids from Human Pluripotent Stem Cells. Methods Mol Biol. 2021; 2389:177-199. DOI: 10.1007/978-1-0716-1783-0_15. View

Zhu L, Stein L, Kim D, Ho K, Yu G, Zhan L . Klotho controls the brain-immune system interface in the choroid plexus. Proc Natl Acad Sci U S A. 2018; 115(48):E11388-E11396. PMC: 6275534. DOI: 10.1073/pnas.1808609115. View

Johansson P, Irmler M, Acampora D, Beckers J, Simeone A, Gotz M . The transcription factor Otx2 regulates choroid plexus development and function. Development. 2013; 140(5):1055-66. DOI: 10.1242/dev.090860. View

Monnot A, Zheng W . Culture of choroid plexus epithelial cells and in vitro model of blood-CSF barrier. Methods Mol Biol. 2012; 945:13-29. PMC: 3982224. DOI: 10.1007/978-1-62703-125-7_2. View

Huang X, Liu J, Ketova T, Fleming J, Grover V, Cooper M . Transventricular delivery of Sonic hedgehog is essential to cerebellar ventricular zone development. Proc Natl Acad Sci U S A. 2010; 107(18):8422-7. PMC: 2889567. DOI: 10.1073/pnas.0911838107. View

Vaughan D, Peters A . Neuroglial cells in the cerebral cortex of rats from young adulthood to old age: an electron microscope study. J Neurocytol. 1974; 3(4):405-29. DOI: 10.1007/BF01098730. View

Pellegrini L, Bonfio C, Chadwick J, Begum F, Skehel M, Lancaster M . Human CNS barrier-forming organoids with cerebrospinal fluid production. Science. 2020; 369(6500). PMC: 7116154. DOI: 10.1126/science.aaz5626. View

Delery E, MacLean A . Culture Model for Non-human Primate Choroid Plexus. Front Cell Neurosci. 2019; 13:396. PMC: 6724611. DOI: 10.3389/fncel.2019.00396. View

10.

Van Hoecke L, Van Cauwenberghe C, Dominko K, Van Imschoot G, Van Wonterghem E, Castelein J . Involvement of the Choroid Plexus in the Pathogenesis of Niemann-Pick Disease Type C. Front Cell Neurosci. 2021; 15:757482. PMC: 8555471. DOI: 10.3389/fncel.2021.757482. View

11.

Dani N, Herbst R, McCabe C, Green G, Kaiser K, Head J . A cellular and spatial map of the choroid plexus across brain ventricles and ages. Cell. 2021; 184(11):3056-3074.e21. PMC: 8214809. DOI: 10.1016/j.cell.2021.04.003. View

12.

Iram T, Kern F, Kaur A, Myneni S, Morningstar A, Shin H . Young CSF restores oligodendrogenesis and memory in aged mice via Fgf17. Nature. 2022; 605(7910):509-515. PMC: 9377328. DOI: 10.1038/s41586-022-04722-0. View

13.

Revah O, Gore F, Kelley K, Andersen J, Sakai N, Chen X . Maturation and circuit integration of transplanted human cortical organoids. Nature. 2022; 610(7931):319-326. PMC: 9556304. DOI: 10.1038/s41586-022-05277-w. View

14.

Pham M, Pollock K, Rose M, Cary W, Stewart H, Zhou P . Generation of human vascularized brain organoids. Neuroreport. 2018; 29(7):588-593. PMC: 6476536. DOI: 10.1097/WNR.0000000000001014. View

15.

Cakir B, Xiang Y, Tanaka Y, Kural M, Parent M, Kang Y . Engineering of human brain organoids with a functional vascular-like system. Nat Methods. 2019; 16(11):1169-1175. PMC: 6918722. DOI: 10.1038/s41592-019-0586-5. View

16.

Saunders N, Dziegielewska K, Fame R, Lehtinen M, Liddelow S . The choroid plexus: a missing link in our understanding of brain development and function. Physiol Rev. 2022; 103(1):919-956. PMC: 9678431. DOI: 10.1152/physrev.00060.2021. View

17.

AlFatah Mansour A, Goncalves J, Bloyd C, Li H, Fernandes S, Quang D . An in vivo model of functional and vascularized human brain organoids. Nat Biotechnol. 2018; 36(5):432-441. PMC: 6331203. DOI: 10.1038/nbt.4127. View