STEMdiff™ Choroid Plexus Organoid Differentiation Kit

Culture medium kit for establishment of organoid models containing human pluripotent stem cell-derived choroid plexus-like epithelium

STEMdiff™ Choroid Plexus Organoid Differentiation Kit

Culture medium kit for establishment of organoid models containing human pluripotent stem cell-derived choroid plexus-like epithelium

STEMdiff Choroid Plexus Organoid Differentiation Kit
1 Kit
399 USD
Catalog # 100-0824

Culture medium kit for establishment and maturation of human pluripotent stem cell-derived choroid plexus organoids

Product Advantages


  • Adopt a relevant human CNS barrier model for screening and modeling applications with the precise control of an in vitro system

  • Eliminate the need for complex co-cultures or Transwell® systems in your screening pipeline

  • Extract CSF to identify human CNS-specific biomarkers in a 100% blood-free system, without requiring a lumbar puncture

  • Obtain 50X more CSF from a single kit than obtained from a single mouse, and eliminate the donor-to-donor variability of pooled clinical samples

What's Included

  • STEMdiff™ Neural Organoid Basal Medium 3, 100 mL
  • STEMdiff™ Neural Organoid Basal Medium 4, 250 mL
  • STEMdiff™ Neural Organoid Supplement E, 10 mL
  • STEMdiff™ Neural Organoid Supplement F, 0.5 mL
  • STEMdiff™ Neural Organoid Supplement G, 0.25 mL
  • STEMdiff™ Neural Organoid Supplement H, 0.5 mL
  • STEMdiff™ Neural Organoid Supplement I, 4.5 mL
  • STEMdiff™ Neural Organoid Supplement J, 0.5 mL
Products for Your Protocol
To see all required products for your protocol, please consult the Product Information Sheet.

Overview

Take an in vitro approach to human neural biomarker discovery and central nervous system (CNS) permeability with human pluripotent stem cell (hPSC)-derived organoids patterned to the choroid plexus, the specialized brain epithelium that forms the blood-cerebrospinal fluid barrier. With a three-dimensional tissue structure and functional barrier epithelium, you’ll achieve unprecedented access and control in studying CNS permeability for drug distribution studies, as well as for production and extraction of cerebrospinal fluid (CSF).

Using STEMdiff™ Choroid Plexus Differentiation Kit’s simple five-stage protocol, you can efficiently and reproducibly generate choroid plexus organoids for a variety of downstream applications. The kit is based on the formulation published by Pellegrini et al. (Science, 2020), which was awarded a 3Rs Prize for its breakthroughs in animal reduction, replacement, and refinement. Choroid plexus organoid formation is initiated through an intermediate embryoid body (EB) formation step followed by expansion of neuroepithelia and patterning to choroid plexus-like epithelium. After a maturation period, organoids generated using this kit feature cystic structures filled with a fluid resembling CSF and surrounded by an epithelial layer expressing ependymal choroid plexus-specific markers (TTR, CLIC6, AQP1).

For extended periods of organoid culture (> 40 days), the components required for organoid maturation can be purchased as STEMdiff™ Choroid Plexus Organoid Maturation Kit (Catalog #100-0825).
Subtype
Specialized Media
Cell Type
Epithelial Cells, PSC-Derived, Neural Cells, PSC-Derived, Neural Stem and Progenitor Cells, Pluripotent Stem Cells
Species
Human
Application
Cell Culture, Differentiation, Functional Assay, Organoid Culture, Phenotyping, Toxicity Assay
Brand
STEMdiff
Area of Interest
Disease Modeling, Drug Discovery and Toxicity Testing, Epithelial Cell Biology, Extracellular Vesicle Research, Neuroscience, Organoids
Formulation
Serum-Free

Data Figures

Cell culture protocol diagram to generate hPSC-derived choroid plexus organoids

Figure 1. Protocol Schematic for the STEMdiff™ Choroid Plexus Organoid Differentiation and Maturation Kits

Choroid plexus organoids can be generated from human embryonic stem (ES) or induced pluripotent stem (iPS) cells in 30 days. The protocol begins with embryoid body (EB) formation, followed by expansion of neuroepithelia and patterning to choroid plexus-like epithelium. After a period of epithelial maturation including extensive bubbling, the organoids develop cystic structures surrounded by an ependymal epithelial layer and filled with a fluid resembling cerebrospinal fluid (CSF).

Brightfield phase-contrast images of whole choroid plexus organoid and of cuboidal epithelial morphology

Figure 2. Organoids Generated Using the STEMdiff™ Choroid Plexus Organoid Differentiation and Maturation Kits Exhibit Fluid-Filled Cysts and Ependymal Epithelial Morphology

(A) Phase-contrast image of a whole choroid plexus organoid at day 40 using the STEMdiff™ Choroid Plexus Organoid Differentiation and Maturation Kits. Organoids exhibit translucent cysts containing a CSF-like fluid. (B) Epithelia on choroid plexus organoids exhibit a tightly packed and cuboidal morphology typical of ependymal cells that line the brain ventricles in vivo.

Immunostained choroid plexus organoids showing markers of ependymal epithelium TTR and CLIC6, with separate bundles of MAP2-positive neurons

Figure 3. Choroid Plexus Organoids Express Markers of Ependymal Epithelium and Show Limited Expression of Neuroepithelial Markers

Immunohistological analysis on whole cleared and stained (Masselink et al. 2019) choroid plexus organoids at day 40 reveals that their cystic structures are (A) TTR- (green) and (B) CLIC6- (red) positive. (C) These ependymal cell markers are highly co-localized. (D) Bundles of MAP2-positive (magenta) neurons are observed but are not co-localized with the ependymal cell markers.

Western blot with 2 columns showing a band in extracted CSF between the 37 and 50 kDa molecular weight markers, but not in spent culture media

Figure 4. Fluid Extracted from Cysts in Choroid Plexus Organoids is Enriched with Clusterin Protein, A Marker of Cerebrospinal Fluid (CSF)

CSF-like fluid was extracted from cysts contained in day 40 choroid plexus organoids using a 28G syringe. A western blot was performed on the extracted fluid to detect Clusterin and shows a band between the 37 and 50 kDa molecular weight marker. Clusterin is a soluble secreted chaperone protein found in high abundance in CSF.

Protocols and Documentation

Find supporting information and directions for use in the Product Information Sheet or explore additional protocols below.

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Safety Data Sheet 1
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Safety Data Sheet 8
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Applications

This product is designed for use in the following research area(s) as part of the highlighted workflow stage(s). Explore these workflows to learn more about the other products we offer to support each research area.

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