Development, Characterization and Potential Applications of a Multicellular Spheroidal Human Blood–Brain Barrier Model Integrating Three Conditionally Immortalized Cell Lines

Kitamura, Kazuko; Umehara, Kenta; Ito, Ryo; Yamaura, Yoshiyuki; Komori, Takafumi; Morio, Hanae; Akita, Hidetaka; Furihata, Tomomi

doi:10.1248/bpb.b21-00218

Cited by 14 publications

(12 citation statements)

References 35 publications

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“…The hiMCS-BBB1.0 and 2.0 models were prepared for immunocytochemistry (ICC). Approximately 10–20 spheroids were pooled in a 1.5 mL Protein LoBind tube and washed with PBS(−), after which ICC was performed as described previously . The primary and secondary antibodies used are listed in Table S1.…”

Section: Methodsmentioning

confidence: 99%

“…The spheroid models were developed in two different protocols. Protocol 1 is the same as described in our previous report 18 in which HASTR/ci35 and HBPC/ci37 cells were simultaneously seeded on the first day and cultured for 48 h, after which HBMEC/ci18 cells were added into the well and incubated for another 48 h (Figure 1A). The resulting model is hereafter referred to as the hiMCS-BBB1.0 model.…”

Section: ■ Materials and Methodsmentioning

confidence: 99%

“…While conventional two-dimensional (transwell) BBB models have been used in BBB permeability studies of various drugs, more physiologically relevant BBB models with better functionality will be unequivocally preferable. To date, fabricated three-dimensional (3D) culture methods, such as organ-on-a-chip devices and spheroids/organoids, have emerged as new ways to build BBB models. − As a part of such research efforts, we have recently employed a layered spheroid culture method to establish a human immortalized cell-based multicellular spheroidal BBB model (hiMCS-BBB model), where the immortalized BMEC forms an outer layer over an internal core made of immortalized pericytes and immortalized astrocytes to create a BBB. As a result of this in vivo BBB-like structure, our hiMCS-BBB models express intercellular junction proteins ( e.g.…”

Section: Introductionmentioning

confidence: 99%

“…To date, fabricated three-dimensional (3D) culture methods, such as organ-on-a-chip devices and spheroids/ organoids, have emerged as new ways to build BBB models. 13−17 As a part of such research efforts, we have recently employed a layered spheroid culture method to establish a human immortalized cell-based multicellular spheroidal BBB model (hiMCS-BBB model), 18 where the immortalized BMEC forms an outer layer over an internal core made of immortalized pericytes and immortalized astrocytes to create a BBB. As a result of this in vivo BBB-like structure, our hiMCS-BBB models express intercellular junction proteins (e.g., claudin-5 and zonula occludens-1 [ZO-1]) and efflux transporters (e.g., P-gp and BCRP) at significantly higher levels than those observed in corresponding Transwell system-based BBB models.…”

Section: ■ Introductionmentioning

confidence: 99%

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Human Immortalized Cell-Based Blood–Brain Barrier Spheroid Models Offer an Evaluation Tool for the Brain Penetration Properties of Macromolecules

et al. 2022

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Blood–brain barrier (BBB)-permeable middle- or macromolecules (middle/macromolecules) have recently attracted significant attention as new drug delivery carriers into the human brain via receptor-mediated transcytosis (RMT). During the development process of such carriers, it is necessary to thoroughly evaluate their human BBB permeability levels. In such evaluations, our recently established human immortalized cell-based multicellular spheroidal BBB models (hiMCS-BBB models) have shown high potential. However, the specifics of those capabilities have yet to be elucidated. Therefore, in this study, we characterize the ability of the hiMCS-BBB models to evaluate RMT-mediated BBB penetration properties of middle/macromolecules. More specifically, we began by validating transferrin receptor (TfR)-mediated RMT functionalities using transferrin in the hiMCS-BBB models and then examined the BBB permeability levels of MEM189 antibodies (known BBB-permeable anti-TfR antibodies). The obtained results showed that, as with the case of transferrin, temperature-dependent uptake of MEM189 antibodies was observed in the hiMCS-BBB models, and the extent of that uptake increased in a time-dependent manner until reaching a plateau after around 2 h. To further expand the evaluation applicability of the models, we also examined the BBB permeability levels of the recently developed SLS cyclic peptide and observed that peptide uptake was also temperature-dependent. To summarize, our results show that the hiMCS-BBB models possess the ability to evaluate the RMT-mediated BBB-permeable properties of antibodies and peptides and thus have the potential to provide valuable tools for use in the exploration and identification of middle/macromolecules showing excellent BBB permeability levels, thereby contributing powerfully to the development of new drug delivery carriers for transporting drugs into the human brain.

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Section: Methodsmentioning

confidence: 99%

Section: ■ Materials and Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: ■ Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Human Immortalized Cell-Based Blood–Brain Barrier Spheroid Models Offer an Evaluation Tool for the Brain Penetration Properties of Macromolecules

et al. 2022

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“…Up to six different cell types (BMECs, astrocytes, pericytes, microglia cells, oligodendrocytes, and neurons) [ 101 , 102 , 103 ]…”

Section: Figurementioning

confidence: 99%

In Vitro Modeling of the Blood–Brain Barrier for the Study of Physiological Conditions and Alzheimer’s Disease

et al. 2022

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The blood–brain barrier (BBB) is an essential structure for the maintenance of brain homeostasis. Alterations to the BBB are linked with a myriad of pathological conditions and play a significant role in the onset and evolution of neurodegenerative diseases, including Alzheimer’s disease. Thus, a deeper understanding of the BBB’s structure and function is mandatory for a better knowledge of neurodegenerative disorders and the development of effective therapies. Because studying the BBB in vivo imposes overwhelming difficulties, the in vitro approach remains the main possible way of research. With many in vitro BBB models having been developed over the last years, the main aim of this review is to systematically present the most relevant designs used in neurological research. In the first part of the article, the physiological and structural–functional parameters of the human BBB are detailed. Subsequently, available BBB models are presented in a comparative approach, highlighting their advantages and limitations. Finally, the new perspectives related to the study of Alzheimer’s disease with the help of novel devices that mimic the in vivo human BBB milieu gives the paper significant originality.

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An overview of in vitro 3D models of the blood-brain barrier as a tool to predict the in vivo permeability of nanomedicines

Pérez-López

Torres-Suárez

Aparicio-Blanco

2023

Advanced Drug Delivery Reviews

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Development, Characterization and Potential Applications of a Multicellular Spheroidal Human Blood–Brain Barrier Model Integrating Three Conditionally Immortalized Cell Lines

Cited by 14 publications

References 35 publications

Human Immortalized Cell-Based Blood–Brain Barrier Spheroid Models Offer an Evaluation Tool for the Brain Penetration Properties of Macromolecules

Human Immortalized Cell-Based Blood–Brain Barrier Spheroid Models Offer an Evaluation Tool for the Brain Penetration Properties of Macromolecules

In Vitro Modeling of the Blood–Brain Barrier for the Study of Physiological Conditions and Alzheimer’s Disease

An overview of in vitro 3D models of the blood-brain barrier as a tool to predict the in vivo permeability of nanomedicines

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