Limitations of the hCMEC/D3 cell line as a model for Aβ clearance by the human blood‐brain barrier

Biemans, Elisanne A.L.M.; Jäkel, Lieke; Waal, R.M.W. de; Kuiperij, H. Bea; Verbeek, Marcel M.

doi:10.1002/jnr.23964

Cited by 55 publications

(36 citation statements)

References 42 publications

(60 reference statements)

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…We observed much lower TEER values when we used inserts with 0.4-µm pores (not shown). These results were similar to what was found in a recent study, although even those barriers were reported to still maintain integrity with respect to large molecules ( 65 ).…”

Section: Methodssupporting

confidence: 92%

Trojan Horse Transit Contributes to Blood-Brain Barrier Crossing of a Eukaryotic Pathogen

Santiago-Tirado

Onken

Cooper

et al. 2017

mBio

182

196

View full text Add to dashboard Cite

The blood-brain barrier (BBB) protects the central nervous system (CNS) by restricting the passage of molecules and microorganisms. Despite this barrier, however, the fungal pathogen Cryptococcus neoformans invades the brain, causing a meningoencephalitis that is estimated to kill over 600,000 people annually. Cryptococcal infection begins in the lung, and experimental evidence suggests that host phagocytes play a role in subsequent dissemination, although this role remains ill defined. Additionally, the disparate experimental approaches that have been used to probe various potential routes of BBB transit make it impossible to assess their relative contributions, confounding any integrated understanding of cryptococcal brain entry. Here we used an in vitro model BBB to show that a “Trojan horse” mechanism contributes significantly to fungal barrier crossing and that host factors regulate this process independently of free fungal transit. We also, for the first time, directly imaged C. neoformans-containing phagocytes crossing the BBB, showing that they do so via transendothelial pores. Finally, we found that Trojan horse crossing enables CNS entry of fungal mutants that cannot otherwise traverse the BBB, and we demonstrate additional intercellular interactions that may contribute to brain entry. Our work elucidates the mechanism of cryptococcal brain invasion and offers approaches to study other neuropathogens.

show abstract

Section: Methodssupporting

confidence: 92%

Trojan Horse Transit Contributes to Blood-Brain Barrier Crossing of a Eukaryotic Pathogen

Santiago-Tirado

Onken

Cooper

et al. 2017

mBio

182

196

View full text Add to dashboard Cite

show abstract

“…In comparison, the model cultivated with standard medium allowed the maximal TEER value after19 days and was stable only during 2 days. Similarly, the other models of human BBB found in the literature, show a short stability, about 2 days only (Biemans et al, 2017;Eigenmann et al, 2016bEigenmann et al, , 2013Weksler et al, 2013). We also observed that HBEC-5i cells with HA conditioned medium exhibited a better permeability for Na-Fl than the hCMEC/D3 cells commonly used in the literature (5.7 x 10 -6 cm.s -1 and 13 x 10 -6 cm.s -1 respectively) (Eigenmann et al, 2013).…”

Section: Discussionsupporting

confidence: 83%

Assessment of HBEC-5i endothelial cell line cultivated in astrocyte conditioned medium as a human blood-brain barrier model for ABC drug transport studies

Puech

Hodin

Forest

et al. 2018

International Journal of Pharmaceutics

View full text Add to dashboard Cite

Endothelial cells are main components of the Blood-Brain Barrier (BBB) and form a tight monolayer that regulates the passage of molecules, with the ATP-Binding Cassette (ABC) transporters efflux pumps. We have developed a human in vitro model of HBEC-5i endothelial cells cultivated alone or with human astrocytes conditioned medium on insert. HBEC-5i cells showed a tight monolayer within 14 days, expressing ZO-1 and claudin 5, a low apparent permeability to small molecules, with a TEER stability during five days. The P-gp, BCRP, MRPs transporters were well expressed and functional. Accumulation and efflux ratio measurement with different ABC transporters substrates (Rhodamine 123, BCECF AM, Hoechst 33342) and inhibitors (verapamil, Ko143, probenecid and cyclosporin A) were conducted. At barrier level, the functionality of ABC transporters was three-fold enhanced in astrocyte conditioned medium. We validated our model by the transport of pharmacological substrates: caffeine, rivaroxaban, and methotrexate. The rivaroxaban and methotrexate were released with an efflux ratio>3 and were decreased by more than half with inhibitors. HBEC-5i model could be used as relevant tool in preclinical studies for assessing the permeability of therapeutic molecules to cross human BBB.

show abstract

“…The BBB prevents virus circulating in blood from entering the brain. The human cerebral microvascular endothelial cell line (hCMEC/D3) is a stable, easily grown BBB cellular model used in a wide range of research areas, including passage of infectious micro-organisms across the BBB [15][16][17][18][19]. There are reports showing that hCMEC/D3 cells are susceptible to ZIKV infection, leading to the speculation that ZIKV has the ability to cross the BBB [18,20].…”

Section: Introductionmentioning

confidence: 99%

Basic insights into Zika virus infection of neuroglial and brain endothelial cells

Mutso

John

Ling

et al. 2020

Journal of General Virology

View full text Add to dashboard Cite

Zika virus (ZIKV) has recently emerged as an important human pathogen due to the strong evidence that it causes disease of the central nervous system, particularly microcephaly and Guillain–Barré syndrome. The pathogenesis of disease, including mechanisms of neuroinvasion, may include both invasion via the blood–brain barrier and via peripheral (including cranial) nerves. Cellular responses to infection are also poorly understood. This study characterizes the in vitro infection of laboratory-adapted ZIKV African MR766 and two Asian strains of (1) brain endothelial cells (hCMEC/D3 cell line) and (2) olfactory ensheathing cells (OECs) (the neuroglia populating cranial nerve I and the olfactory bulb; both human and mouse OEC lines) in comparison to kidney epithelial cells (Vero cells, in which ZIKV infection is well characterized). Readouts included infection kinetics, intracellular virus localization, viral persistence and cytokine responses. Although not as high as in Vero cells, viral titres exceeded 104 plaque-forming units (p.f.u.) ml−1 in the endothelial/neuroglial cell types, except hOECs. Despite these substantial titres, a relatively small proportion of neuroglial cells were primarily infected. Immunolabelling of infected cells revealed localization of the ZIKV envelope and NS3 proteins in the cytoplasm; NS3 staining overlapped with that of dsRNA replication intermediate and the endoplasmic reticulum (ER). Infected OECs and endothelial cells produced high levels of pro-inflammatory chemokines. Nevertheless, ZIKV was also able to establish persistent infection in hOEC and hCMEC/D3 cells. Taken together, these results provide basic insights into ZIKV infection of endothelial and neuroglial cells and will form the basis for further study of ZIKV disease mechanisms.

show abstract

Limitations of the hCMEC/D3 cell line as a model for Aβ clearance by the human blood‐brain barrier

Cited by 55 publications

References 42 publications

Trojan Horse Transit Contributes to Blood-Brain Barrier Crossing of a Eukaryotic Pathogen

Trojan Horse Transit Contributes to Blood-Brain Barrier Crossing of a Eukaryotic Pathogen

Assessment of HBEC-5i endothelial cell line cultivated in astrocyte conditioned medium as a human blood-brain barrier model for ABC drug transport studies

Basic insights into Zika virus infection of neuroglial and brain endothelial cells

Contact Info

Product

Resources

About