A subscription to JoVE is required to view this content. Sign in or start your free trial.

Method Article

An Advanced Co-Culture Model of the Human Blood-Cerebrospinal Fluid Barrier for Separate Analysis of Choroid Plexus Epithelium and Endothelium

415 views

⸱

DOI:

10.3791/69275

⸱

November 28th, 2025

 ,  ,  ,  ,  ,  , 
1Pediatric Infectious Diseases, Department of Pediatrics, Medical Faculty Mannheim, Heidelberg University, 2European Center for Angioscience, Medical Faculty Mannheim, Heidelberg University, 3Department of Medical Statistics, Biomathematics and Information Processing, Medical Faculty Mannheim, Heidelberg University, 4Laboratory of Clinical Regenerative Medicine, Department of Neurosurgery, Faculty of Medicine, University of Tsukuba

In This Article

Summary

A co-culture of human epithelial and endothelial cells of the choroid plexus was established to generate an advanced in vitro model of the blood-cerebrospinal fluid barrier. We describe the separate isolation of epithelium and endothelium for subsequent RNA preparation and transcriptional analysis under selected conditions.

Abstract

The highly perfused choroid plexus (CP), located in the ventricular system of the brain, consists of several structural components, including a cuboid epithelium and a fenestrated endothelium, and is the location of the blood-cerebrospinal fluid barrier (BCSFB). To investigate the CP in vitro, several different cell culture setups are conceivable. An inverted cell culture filter model with strong barrier function, based on human epithelial CP papilloma (HIBCPP) cells, was established by our laboratory and is used extensively. Recently, we were able to generate a stable immortalized human CP endothelial cell line (iHCPEnC). Combining both cell lines, we have assembled an advanced functional two-cell-type model that developed an increased barrier function compared to HIBCPP cells alone. Since this model represents both the CP epithelium and endothelium, it allows for the investigation of the interplay between the two cell types under selected conditions, such as infection, inflammation, or others. In this protocol, we describe methods to set up this two-cell-type model with subsequent separation of the epithelial and endothelial cells for transcriptional analyses. We describe PCR primers specific for epithelium and endothelium, which serve to detect possible cross-contamination between the two cell types. To investigate a possible impact of the CP endothelium on the epithelium in this model under inflammatory conditions, the expression levels of selected genes in the context of lipopolysaccharide (LPS) treatment were analyzed in the HIBCPP cell fraction. The advanced CP in vitro model and the possibility to separately analyze epithelial and endothelial responses are of significant interest for research on cellular processes involving biological and pathological functions of the CP.

Introduction

The choroid plexus (CP) is a highly vascularized conglomerate of different populations of cells located in all four ventricles of the brain. Among other functions, it serves as a physiological barrier, the so-called blood-cerebrospinal fluid barrier (BCSFB), between the central nervous system (CNS) and the blood, preventing the brain from, e.g., infectious substances circulating in the blood. The CP consists of different cell types, including the polarized epithelial cells, containing tight- and adherence junctions as well as desmosomes, sitting on a basement membrane. The epithelial cells' surface is enlarged by the presence of microvilli on the luminal surface. Fenestrated endothelial capillaries, surrounded by connective tissue such as fibroblasts and immune cells, are located underneath the basement membrane1,2,3. It was recently reported that CP endothelial cells are able to enhance BCSFB integrity together with the epithelial cells and thus might play a role in brain protection during inflammatory processes4,5.

Up to now, the inverted culture of human epithelial CP papilloma (HIBCPP) cells is the most frequently used in vitro model to mimic a human BCSFB with high transepithelial electrical resistance (TEER) values and a strong barrier function. To gain a better understanding of cellular functions and avoid animal experiments at the same time it is important to get as much information out of in vitro experiments as possible. The first stable immortalized human CP endothelial cell line (iHCPEnC) was established recently in our laboratory6,7. With the help of HIBCPP cells and iHCPEnC we are able to assemble a co-culture in vitro model imitating the BCSFB in a more advanced fashion8. Using this two-cell-type model, we could demonstrate that the co-culture of HIBCPP cells and iHCPEnC leads to increased TEER values compared to HIBCPP cells alone. It is important to note that iHCPEnC alone does not develop a TEER7,8. Separate preparation and subsequent analyses of the two cell types employed in this model would allow us to investigate their interplay under healthy and diseased conditions.

Here we describe two different approaches to achieve this aim. The basis is that, in both cases, the approach is based on the inverted cell-culture model of the HIBCPP cells (Figure 1). Method 1, the "direct co-culture" method, comprises both cells on one filter insert. HIBCPP cells are grown in an inverted fashion prior to seeding iHCPEnC in standard fashion7,8. Method 2, the "membrane-in-insert" culture, here we added a separate membrane with the same properties as the one used in the filter inserts. This additional membrane is put into a filter insert where HIBCPP cells are already growing in inverted culture. iHCPEnC are seeded onto the additional membrane in standard culture fashion.

Subsequent separation of the cells for RNA analysis allows insights into the transcriptional reaction of the CP epithelium and endothelium under selected conditions. We describe PCR primers specific for HIBCPP cells and iHCPEnC that enable the confirmation of the successful and clean separation of both cell types. Following treatment with lipopolysaccharide (LPS), we use the described models to investigate the distinct responses of the CP epithelium in the presence of the endothelium under inflammatory conditions.

Access restricted. Please log in or start a trial to view this content.

Protocol

1. HIBCPP cells

NOTE: In the following, we describe the culture and maintenance of HIBCPP cells, as well as the preparation of inverted cell culture filter inserts with HIBCPP cells. This protocol is designed for the growth of HIBCPP cells on 12-well cell culture filter inserts. A detailed protocol with accompanying video for culture of HIBCPP cells on 24-well cell culture filter inserts has been published previously9.

  1. Taking HIBCPP cells into culture
    1. Thaw a vial of HIBCPP cells at room temperature (RT).
    2. Resuspend the content of the vial with 10 mL of DMEM/HAMS-F12 medium containing 10% fetal calf serum (FCS) and 5 µg Insulin/mL of pre-warmed medium (10% HIBCPP medium).
    3. Put the resuspended HIBCPP cells into a T75 cell culture flask and incubate for 24 h at 37 °C with 5% CO2.
    4. Change the medium of the flask to remove the DMSO 24 h after seeding the cells.
    5. Culture the cells until 90% confluency, this step takes around 2 weeks. Change the medium twice a week.
  2. Continuous culture of HIBCPP cells
    NOTE: For continuous culture, split HIBCPP cells 1:3 into new T75 cell culture flasks.
    1. Remove the culture medium from the T75 flask.
    2. Wash cells once with 10 mL of PBS.
    3. Remove the PBS.
    4. Detach HIBCPP cells with the help of 3 mL of 0.25% Trypsin-EDTA. This step takes around 10-20 min at 37 °C. Swirl the flask in the meantime to ensure the complete covering of the cell layer with Trypsin-EDTA.
    5. Add 7 mL of HIBCPP medium and centrifuge at 50 x g for 10 min at RT.
    6. Resuspend the cell pellet in 30 mL of 10% HIBCPP medium and spread to 3 new T75 flasks.
    7. Culture the cells until 90% confluency; the cells will reach confluency in around 1 week.
    8. Change the medium twice a week.
      NOTE: HIBCPP cells are not growth-inhibited; therefore, the flask will turn confluent, and cells will keep growing and form multilayers.
  3. Seeding HIBCPP cells in inverted culture
    1. Remove the culture medium from the T75 flask.
    2. Wash cells once with 10 mL of PBS.
    3. Remove the PBS.
    4. Detach HIBCPP cells with the help of 3 mL of 0.25% Trypsin-EDTA. This step takes around 10-20 min at 37 °C. Swirl the flask in the meantime to ensure the complete covering of the cell layer with Trypsin-EDTA.
    5. In the meantime, prepare the filter inserts. Turn 12-well filter inserts in an upside-down orientation into a 6-well cell culture plate with the help of forceps and fill them with pre-warmed 10% HIBCPP medium. Pre-wet the membrane with a drop of medium on top of each filter insert.
    6. Resuspend trypsinized cells with 17 mL of 10% HIBCPP medium.
    7. Centrifuge at 50 x g for 10 min at RT and adjust HIBCPP cells to a concentration of 1 × 106 cells/mL.
      NOTE: HIBCPP cells tend to form small agglomerates. It is not necessary to dissolve them completely before seeding onto filter inserts. Ensure 90% of the cells are single.
    8. Seed HIBCPP cells in an inverted fashion on 12-well filter inserts with a density of 4 × 105 cells/insert.
    9. Wait until the cell suspension is taken up by the membrane. Close the lid of the 6-well cell culture plate.
    10. Put the plates with the inverted inserts into the incubator.
    11. 24 h after seeding, turn the inserts with the help of forceps to their correct orientation into a new 12-well cell culture plate. Put 2 mL of 10% HIBCPP medium in the lower compartment and 1 mL into the upper compartment.
    12. Leave the cells for 5 to 6 days without any medium change, and start the determination of TEER and the setting up of co-culture.

2. Determination of TEER

NOTE: Determination of TEER values is performed with the help of a MilliCell ERS2 and a chopstick electrode. A blank measurement from cell culture filter inserts in medium without cells (Ohmblank) is required for calculation.

  1. Store the chopstick electrode dry. Prior to starting the measurement, put the electrode into 80% ethanol for 10 min for disinfection.
  2. Afterwards, rinse the electrode with PBS.
  3. Starting the measurement: Put the long end of the electrode into the well and the short end of the electrode into the filter insert containing the seeded cells.
  4. Record the Ohm value of the 12-well cell culture filter insert (Ohmsample).
  5. Calculate the TEER as follows: Ohm·cm2 = (Ohmsample- Ohmblank) × 1.1 cm2.
  6. After finishing the measurement, rinse the electrode with 80% ethanol and then store dry.
    NOTE: In between measurements of different experimental conditions or measurements of different cell lines, the chopstick electrode needs to be disinfected and rinsed with PBS to prevent cross-contamination.

3. iHCPEnC

  1. Taking iHCPEnC into culture
    1. Thaw a vial of iHCPEnC at RT.
    2. Resuspend the content of the vial with pre-warmed 5 mL of Complete Classic Endothelial Medium (iHCPEnC medium).
    3. Centrifuge at 500 g for 5 min at RT.
    4. In the meantime, coat a T25 cell culture flask with 2 mL of Attachment Factor for approximately 1 min. Aspirate the Attachment Factor and leave the flask until seeding the iHCPEnC.
    5. Resuspend the cell pellet with 5 mL of iHCPEnC medium and put it into the prepared T25 cell culture flask.
    6. Incubate at 37 °C with 5% CO2 for 1 week without any change of medium.
  2. Continuous culture of iHCPEnC
    ​NOTE: For continuous culture, split iHCPEnC 1:4 into new T25 cell culture flasks.
    1. Remove the iHCPEnC medium and wash cells once with 3 mL of PBS. Be careful to avoid detaching of cells.
    2. Aspirate the PBS.
    3. Detach confluent iHCPEnC with the help of 1 mL of Trypsin-EDTA. This step takes around 3-5 min at RT.
    4. Resuspend the cells in 5 mL of iHCPEnC medium.
    5. Centrifuge at 500 g for 5 min at RT.
    6. In the meantime, coat 4 new T25 flasks with 2 mL of Attachment Factor per flask.
    7. Resuspend the cell pellet with 20 mL of iHCPEnC medium.
    8. Distribute 5 mL of the cell suspension to each of the 4 prepared flasks.
    9. Again, culture the cells for 1 week without any change of medium. They will reach confluency in 1week.
      NOTE: iHCPEnC cells are growth-inhibited; therefore, the flask will turn confluent, and the remaining cells will float.

4. Preparations for co-culture experiments

  1. Method 1: For "direct co-culture" (culturing the epithelial and the endothelial cells on both sides of one insert), do not prepare any additional things.
  2. Method 2: For the "membrane-in-insert" method, perform the following steps:
    1. Punch out 12-well cell culture filter insert-sized membranes with the help of a no. 11 perforating iron (corresponding to a diameter of 11 mm) from a separate Din-A-4 membrane.
    2. Put punched-out membranes into a Petri dish.
    3. Sterilize punched-out membranes for 60 min using ultraviolet (UV) light (UV-C, 254 nm).
      NOTE: Punched-out and sterilized 12-well cell culture filter insert-sized membranes can be prepared in advance and stored under sterile conditions.

5. Setting up the co-culture

NOTE: At this stage, the HIBCPP cells are already prepared for the two-cell type in vitro culture model. HIBCPP cells have been grown for 5 to 6 days (compare steps 1.3.8-1.3.12). HIBCPP cells should start developing a TEER (around 20 Ohm·cm2).

  1. For both, Method 1 ("direct co-culture") and Method 2 ("membrane-in-insert"), replace the 10% HIBCPP medium with 1% HIBCPP medium. For this purpose, put 2 mL of 1% HIBCPP medium into a well of a 12-well cell culture plate.
  2. Empty the inverted culture containing filter insert with the help of a pipette and forceps, and put the filter insert in standard orientation into the prepared 12-well.
  3. For Method 2 ("membrane-in-insert"), prepare filter inserts as follows:
    1. Put the punched-out and sterilized membrane into a prepared 12-well filter insert with HIBCPP cells in inverted culture with the help of forceps.
  4. Put the cell culture plates back into the incubator for the trypsinization of the iHCPEnC (described under step 6).

6. Seeding and cultivation of iHCPEnC in co-culture

NOTE: In the following section, the establishment of a co-culture of HIBCPP cells and iHCPEnC on 12-well cell culture filter inserts is described. A detailed protocol for co-culture of HIBCPP cells and iHCPEnC on 24-well cell culture filter inserts has been described previously8. Before detaching iHCPEnC, calculate how many cells are needed to perform the co-culture. One confluent T25 cell culture flask will provide an approximate amount of 5 × 105 cells.

  1. Remove the medium of a confluent T25 flask containing iHCPEnC.
  2. Wash iHCPEnC with 3 mL of PBS. Avoid detaching of the cells.
  3. Remove the PBS.
  4. Detach confluent iHCPEnC with the help of 1 mL of Trypsin-EDTA. This step takes around 3-5 min at RT.
  5. Resuspend the cells in 5 mL/T25 flask with iHCPEnC medium.
  6. Count the cells in a Neubauer chamber.
  7. Centrifuge at 500 × g for 5 min at RT.
  8. Resuspend the cell pellet to a final concentration of 1 × 105 cells/mL in iHCPEnC medium.
  9. Both Method 1 ("direct co-culture") and Method 2 ("membrane in insert" ): seed 700 µL iHCPEnC (7x105 cells) in a 12-well filter insert.
  10. Monitor TEER values of the co-culture and controls (i.e., HIBCPP cell and iHCPEnC only cultures) for the first time 24 h after seeding iHCPEnC and afterwards daily.

7. LPS treatment

NOTE: Co-cultures should present higher TEER values compared to HIBCPP cell-only cultures (suggested factor: 1.5 or higher).

  1. Measure and record TEER values of cell culture inserts set up for experiments.
  2. Prepare a master mix containing 5 µg/mL LPS in 1% HIBCPP medium.
  3. Prepare new 12-well plates containing 2 mL 1% HIBCPP medium per well.
  4. Empty the inserts with the help of forceps and pipetting.
  5. Place inserts into the prepared wells.
  6. Add 1 mL of the LPS master mix or 1% HIBCPP medium, respectively (negative control).
  7. Incubate for 4 h at 37 °C.
  8. Measure and record TEER values before proceeding to section 8.

8. Separate isolation of RNA from HIBCPP cells and iHCPEnC for subsequent transcriptional analysis

NOTE: To achieve two separate cell fractions and avoid cross-contamination from cells cultured in the co-culture model, the cell fractions are carefully scraped off the insert separately. Perform all steps of RNA isolation for each filter insert without interruption (the inserts are not allowed to run dry).

  1. Wash filter inserts once with PBS. For this purpose, prepare 12-well cell culture plates containing 1.5 mL PBS/well.
  2. Empty the cell culture filter inserts with the help of a pipette and forceps. Place the inserts into the PBS-filled 12-well cell culture plate.
  3. Add 1 mL of PBS into the inserts.
  4. To prepare iHCPEnC and HIBCPP cells for Method 1 ("direct co-culture" ), perform the following steps:
    1. Remove the PBS from the insert and place it into an empty and dry 12-well cell culture plate.
    2. With the help of a pipette tip and pipetting up and down of 10-20 µL of PBS, the iHCPEnC are detached from the insert by scraping.
    3. Place this cell fraction in a 1.5 mL tube containing 350 µL of RNA lysis buffer. Pipette up and down to mix the cells properly with the RNA lysis buffer.
    4. After the harvest of the iHCPEnC fraction is completed, turn the insert downside up into the lid of a cell culture plate.
    5. Take HIBCPP cells off the membrane just like the iHCPEnC fraction, and place them in a prepared 1.5 mL tube containing 350 µL of RLT-β-mercaptoethanol.
      NOTE: If multiple co-cultures are prepared for one condition, take 350 µL of RNA lysis buffer per insert and cell fraction. Single prepared cell fractions of one specific condition are consecutively purified on the same column.
  5. To prepare iHCPEnC and HIBCPP cells for Method 2 ("membrane-in-insert"), perform the following steps:
    1. Take out the additional membrane with the help of forceps and put it into a dry lid of a 12-well cell culture plate. Pipette 100 µL of RNA lysis buffer onto the membrane for lysis of iHCPEnC.
    2. Place this 100 µL lysate into a 1.5 mL tube containing 250 µL of RNA lysis buffer and mix well.
    3. Remove the PBS from the filter insert and put it downside up into the lid of a 12-well cell culture plate. Add 100 µL of RNA lysis buffer onto the membrane of the insert and lyse the HIBCPP cell fraction.
    4. Place this 100 µL lysate into a 1.5 mL tube containing 250 µL of RNA lysis buffer.
  6. Perform RNA isolation according to the manufacturer's protocol.
  7. Measure the quantity of RNA using a spectrophotometer.
  8. Perform reverse transcription and qPCR analysis of RNA samples as described by the manufacturer.
    NOTE: As the co-cultures contain less amounts of iHCPEnC compared to HIBCPP cells, several 12-well filter inserts are needed for isolation of sufficient RNA from iHCPEnC, depending on the subsequent analysis. The RNA lysates of the distinct filter inserts need to be pooled. Reverse transcription and qPCR can be performed as described previously6.

Access restricted. Please log in or start a trial to view this content.

Results

In this protocol, we describe the step-by-step generation of a two-cell-type model of the human CP consisting of epithelium (HIBCPP cells) and endothelium (iHCPEnC) on cell culture filter inserts, as shown schematically in Figure 1. In this model system, HIBCPP cells are grown in an inverted fashion on the lower side of the filter, whereas iHCPEnC are seeded either on the upper side of the filter of the insert or on a second filter placed on top of the membrane of the insert.

Access restricted. Please log in or start a trial to view this content.

Discussion

The human CP consists of several different cell types, including a barrier-forming epithelium and an endothelium of the extended vasculature3. Although the endothelial cells of the CP are fenestrated, recent research has demonstrated their contribution to barrier function at the BCSFB4,7. To study the intricate interactions between the endothelium and epithelium in organs as the CP, advanced in vitro models allowing analysis of bo...

Access restricted. Please log in or start a trial to view this content.

Disclosures

The authors declare that they have no conflict of interest.

Materials

List of materials used in this article
NameCompanyCatalog NumberComments
0.25% Trypsin-EDTALife Technologies 25200056
12 well cell culture plate StarLabCC7682-7512
12 well filter inserts (3 µM pores)Greiner Bio One665631
24 well cell culture plate StarLabCC7672-7524
24 well filter insert  (0.4 µM pores)Greiner Bio One662641
24 well filter insert (3 µM pores)Greiner Bio One662631
6 well cell culture plate StarLabCC7682-7506
AffinityScript cDNA Synthesis Kit (cDNA synthesis )Agilent600559
Attachment FactorCell Systems4ZO-500Part of the Complete Classic Endothelial Medium kit.
Brilliant II SYBR Green qPCR Master Mix (real-time PCR )Agilent600828
Chopstick electrodeMilliporeMERSSTX01This product is discontinued.
Complete Classic Endothelial Medium (iHCPEnC medium)Cell Systems4ZO-500Kit, containing medium, supplements and Attachment Factor for coating.
DMEM/F-12 (1:1) w/ phenolred (10% HIBCPP medium)Thermo Fisher31330-038
DMEM/F-12 (1:1) w/o phenolred (1% HIBCPP medium)Thermo Fisher11039-021
Fetal Calf Serum (FCS) (1% and 10% HIBCPP medium)Thermo Fisher10270-106
Insulin (1% and 10% HIBCPP medium)SigmaI9278
Lipopolysaccharide (LPS)SigmaL3024
MilliCell ERS2 MilliporeMERS00002This product is discontinued.
Nanodrop (Spectrophotometer)Thermo ScientificND1000This product is discontinued.
No. 11 perforating ironWürth880223211
PBSLife Technologies 14190169
Primer designing toolNCBIhttps://www.ncbi.nlm.nih.gov/tools/primer-blast/
Primers SigmaVC00021
Qiagen RNeasy microQiagen74004
RLT+b-ME (part of Qiagen Rneasy micro kit)Qiagen74004
T25 cell culture flaskSarstedt AG &Co KG83,39,10,002
T75 cell culture flaskSarstedt AG &Co KG83,39,11,002
TRAKETCH PET 3.0 cc (separate membrane)SabeuSK4580
UV lightCarl RothH469.1

References

  1. Damkier, H. H., Brown, P. D., Praetorius, J. Cerebrospinal fluid secretion by the choroid plexus. Physiol Rev. 93 (4), 1847-1892 (2013).
  2. Schwerk, C., Schroten, H. In vitro models of the choroid plexus and the blood-cerebrospinal fluid barrier: advances, applications, and perspectives. Hum Cell. 37 (5), 1235-1242 (2024).
  3. Ghersi-Egea, J. F., et al. Molecular anatomy and functions of the choroidal blood-cerebrospinal fluid barrier in health and disease. Acta Neuropathol. 135 (3), 337-361 (2018).
  4. Carloni, S., et al. Identification of a choroid plexus vascular barrier closing during intestinal inflammation. Science. 374 (6566), 439-448 (2021).
  5. Carloni, S., Rescigno, M. Unveiling the gut-brain axis:structural and functional analogies between the gut and the choroid plexus vascular and immune barriers. Semin Immunopathol. 44 (6), 869-882 (2022).
  6. Denzer, L., et al. Transcriptome and functional comparison of primary and immortalized endothelial cells of the human choroid plexus at the blood-cerebrospinal fluid barrier. Int J Mol Sci. 26 (4), 1779(2025).
  7. Muranyi, W., et al. Immortalized human choroid plexus endothelial cells enable an advanced endothelial-epithelial two-cell type in vitro model of the choroid plexus. iScience. 25 (6), 104383(2022).
  8. Herold, R., et al. Protocol for the setup and use of a human choroid plexus endothelial-epithelial two-cell-type in vitro model. STAR Protoc. 3 (4), 101816(2022).
  9. Dinner, S., et al. A choroid plexus epithelial cell-based model of the human blood-cerebrospinal fluid barrier to study bacterial infection from the basolateral side. J Vis Exp. (111), e54061(2016).
  10. Herold, R., et al. Capsule-dependent impact of MAPK signalling on host cell invasion and immune response during infection of the choroid plexus epithelium by Neisseria meningitidis. Fluids Barriers CNS. 18 (1), 53(2021).
  11. Bernd, A., et al. Characterization of efflux transport proteins of the human choroid plexus papilloma cell line HIBCPP, a functional in vitro model of the blood-cerebrospinal fluid barrier. Pharm Res. 32 (9), 2973-2982 (2015).
  12. Dabbagh, F., Schroten, H., Schwerk, C. In vitro models of the blood-cerebrospinal fluid barrier and their applications in the development and research of (neuro)pharmaceuticals. Pharmaceutics. 14 (8), 1729(2022).
  13. Lun, M. P., Monuki, E. S., Lehtinen, M. K. Development and functions of the choroid plexus-cerebrospinal fluid system. Nat Rev Neurosci. 16 (8), 445-457 (2015).
  14. Zhang, W. Y., Liu, Y., Zhang, H. Extracellular matrix: an important regulator of cell functions and skeletal muscle development. Cell Biosci. 11 (1), 65(2021).
  15. Clark, A. J., et al. Functional imaging in microfluidic chambers reveals sensory neuron sensitivity is differentially regulated between neuronal regions. Pain. 159 (7), 1413-1425 (2018).
  16. Kim, J., Koo, B. K., Knoblich, J. A. Human organoids: model systems for human biology and medicine. Nat Rev Mol Cell Biol. 21 (10), 571-584 (2020).
  17. Augustin, H. G., Koh, G. Y. Organotypic vasculature: from descriptive heterogeneity to functional pathophysiology. Science. 357 (6353), eaal2379(2017).
  18. Furtado, A., et al. Circadian rhythmicity of amyloid-beta-related molecules is disrupted in the choroid plexus of a female Alzheimer's disease mouse model. J Neurosci Res. 101 (4), 524-540 (2023).
  19. Borkowski, J. Neisseria meningitidis elicits a pro-inflammatory response involving IκBζ in a human blood-cerebrospinal fluid barrier model. J Neuroinflammation. 11, 163(2014).
  20. Wiatr, M., et al. Distinct migratory pattern of naive and effector T cells through the blood-CSF barrier following Echovirus 30 infection. J Neuroinflammation. 16 (1), 232(2019).
  21. Speidel, A., et al. Transmigration of Trypanosoma brucei across an in vitro blood-cerebrospinal fluid barrier. iScience. 25 (4), 104014(2022).
  22. Haas, J., et al. The choroid plexus is permissive for a preactivated antigen-experienced memory B-cell subset in multiple sclerosis. Front Immunol. 11, 618544(2020).
  23. Nishihara, H., et al. Human CD4+ T cell subsets differ in their abilities to cross endothelial and epithelial brain barriers in vitro. Fluids Barriers CNS. 17 (1), 3(2020).
  24. Erb, U., Schwerk, C., Schroten, H., Karremann, M. Review of functional in vitro models of the blood-cerebrospinal fluid barrier in leukaemia research. J Neurosci Methods. 329, 108478(2020).
  25. Ho, K. H., et al. Activation of Wnt/β-catenin signaling is critical for the tumorigenesis of choroid plexus. Neuro Oncol. 27 (1), 106-122 (2025).
  26. Wagenaars, F., et al. Characterization of thyroid hormone transport in a human choroid plexus papilloma cell line (HIBCPP) as an in vitro blood-cerebrospinal fluid barrier model. Mol Cell Endocrinol. 597, 112449(2025).
  27. Di Tommaso, N., Santopaolo, F., Gasbarrini, A., Ponziani, F. R. The gut-vascular barrier as a new protagonist in intestinal and extraintestinal diseases. Int J Mol Sci. 24 (2), 1470(2023).

Access restricted. Please log in or start a trial to view this content.

Reprints and Permissions

Tags

Choroid Plexus EndotheliumHIBCPP CellsImmortalized Endothelial CellsBarrier FunctionIn Vitro ModelGene Expression AnalysisLipopolysaccharide Treatment
Contact Us
Recommend to Library

Privacy

Terms of Use

Policies

Copyright © 2026 MyJoVE Corporation. All rights reserved.