Drug Delivery Characteristics of the Progenitor Bronchial Epithelial Cell Line VA10

Benediktsdóttir, Berglind Eva; Arason, Ari Jón; Halldórsson, Skarphéðinn; Guðjónsson, Þórarinn; Másson, Már; Baldursson, Ólafur

doi:10.1007/s11095-012-0919-x

Cited by 12 publications

(15 citation statements)

References 47 publications

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“…The ciliated cells covered 10 to 15% of the tissue models' surface. 1 We investigated if the chosen 3D scaffold and addition of primary human airway fibroblasts could improve the mucociliary phenotype. However, when seeded on the SIS, VA10 lacked kinocilia, built an epithelial layer containing microvilli and tight junctions, CK5-positive basal, and CK18-positive differentiated cells, and showed a qualitatively low amount of the mucins 5AC and 5B.…”

Section: Discussionmentioning

confidence: 99%

“…For example, it was reported that kinocilia of the VA10 cell line covered up to 15% of the tissue model's surface area, exhibiting a beating frequency of 6-7 Hz when seeded on transwell inserts and cultured under air-lift conditions. 1 The cell line HBEC3-KT that was seeded on fibroblast-loaded collagen gels developed kinocilia; however, there is only little information on ciliary functionality. 11 To investigate distinct research topics using 3D respiratory epithelial/mucosal tissue models, such as host-pathogen interaction of the respiratory epithelium with Bordetella pertussis that requires the presence of kinocilia for adherence 12 or ciliopathies, for example, primary ciliary dyskinesia (PCD), 13 functional kinocilia and, thus, mucociliary transport are mandatory.…”

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confidence: 99%

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Investigation on Ciliary Functionality of Different Airway Epithelial Cell Lines in Three-Dimensional Cell Culture

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Perniss

et al. 2020

Tissue Engineering Part A

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Three-dimensional respiratory tissue models have been generated using, for example, human primary airway epithelial cells (hAEC) or respective cell lines. To investigate ciliopathies, such as primary ciliary dyskinesia, the presence of functional kinocilia in vitro is an essential prerequisite. Since access to hAEC of healthy donors is limited, we aimed to identify a respiratory epithelial cell line that is capable to display functional kinocilia on at least 60% of the apical surface. Thus, we cultured four different human respiratory cell lines with human primary airway fibroblasts under airlift conditions, characterized the morphology, and analyzed ciliary function. Only one of the tested cell lines showed beating kinocilia; however, <10% of the whole surface was covered and ciliary beating was undirected. Positive control tissue models using hAEC and fibroblasts displayed expected directed ciliary beating pattern around 11 Hz. Our data show that the available cell lines are not suitable for basic and applied research questions whenever functional kinocilia are required and that, rather, hAEC-or human induced pluripotent stem cell-derived tissue models need to be generated.To study ciliopathies or Bordetella pertussis infection in vitro, three-dimensional respiratory tissue models with functional kinocilia covering at least 60% of the model's surface are mandatory. We cultured four respiratory cell lines on a fibroblastloaded biological scaffold and showed that none of them met this requirement. In contrast, primary airway cell-derived models sufficiently reflected the mucociliary phenotype. To further search for an alternative to primary respiratory cells, investigations on other cell lines should be conducted or even new cell lines have to be generated. IntroductionT hree-dimensional (3D) cell cultures of the respiratory epithelium/mucosa afford research on-for examplethe impact of airborne pollutants, host-pathogen interactions, and drug permeation. 1-5 As source for respiratory epithelial cells, primary cells, induced pluripotent stem cells (iPSC), or cell lines can be used. Both primary cells and iPSC provide the opportunity to generate personalized tissue models, for example, to study individual drug responses or drug efficacy. Moreover, they show a high in vitro-in vivo correlation. These models feature a pseudostratified epithelial morphology, barrier properties, basal cells, mucus-producing goblet cells, and ciliated cells facilitating mucociliary clearance. [6][7][8][9]

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Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

Investigation on Ciliary Functionality of Different Airway Epithelial Cell Lines in Three-Dimensional Cell Culture

Lodes

Seidensticker

Perniss

et al. 2020

Tissue Engineering Part A

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“…Indeed, highly differentiated primary human airway epithelial cell layers and cultured HBE-2 bronchial epithelial cells were recently used for infection with B. pertussis to study the role of fimbriae in bacterial adherence to ciliated cells (46). Here we used VA10 epithelial cells that were polarized and differentiated at the air-liquid interface (ALI) and that form pseudostratified epithelial layers with apicobasal polarity, functional tight junctions, and TEER, the hallmarks of epithelial barrier function (47,48). A previous study from our group showed that infection by the opportunistic pathogen Pseudomonas aeruginosa provokes the complete loss of TEER, where the barrier function of VA10 cell layers grown at the ALI is obliterated by a coordinated action of numerous secreted cytotoxic factors (37).…”

Section: Discussionmentioning

confidence: 99%

Bordetella pertussis Adenylate Cyclase Toxin Disrupts Functional Integrity of Bronchial Epithelial Layers

et al. 2018

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The airway epithelium restricts the penetration of inhaled pathogens into the underlying tissue and plays a crucial role in the innate immune defense against respiratory infections. The whooping cough agent, , adheres to ciliated cells of the human airway epithelium and subverts its defense functions through the action of secreted toxins and other virulence factors. We examined the impact of infection and of adenylate cyclase toxin-hemolysin (CyaA) action on the functional integrity of human bronchial epithelial cells cultured at the air-liquid interface (ALI). adhesion to the apical surface of polarized pseudostratified VA10 cell layers provoked a disruption of tight junctions and caused a drop in transepithelial electrical resistance (TEER). The reduction of TEER depended on the capacity of the secreted CyaA toxin to elicit cAMP signaling in epithelial cells through its adenylyl cyclase enzyme activity. Both purified CyaA and cAMP-signaling drugs triggered a decrease in the TEER of VA10 cell layers. Toxin-produced cAMP signaling caused actin cytoskeleton rearrangement and induced mucin 5AC production and interleukin-6 (IL-6) secretion, while it inhibited the IL-17A-induced secretion of the IL-8 chemokine and of the antimicrobial peptide beta-defensin 2. These results indicate that CyaA toxin activity compromises the barrier and innate immune functions ofinfected airway epithelia.

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“…For example, BEAS-2B cells, which were developed by immortalization of normal human bronchial epithelial cells using an adenovirus-SV40 hybrid virus, retain the capacity for squamous differentiation, while 16HBE14o- cells (bronchial epithelium immortalized with plasmid containing a replication deficient SV40 virus genome) can differentiate into ciliated cells in ALI culture [35,41,57]. Similarly, immortalizing bronchial epithelial cells using HPV-16 E6 and E7 alone (VA10 cells) or together with hTERT (NuLi 1–2) resulted in cell lines capable of differentiating into ciliated (VA10) or ciliated and goblet cells (NuLi-2) on ALI culture [39,40,58]. Combined expression of SV40 early antigen and hTERT resulted in immortalization of small airway epithelial cells to generate the cell line SA (SV40 ER + hTERT) which, when injected subcutaneously into a nude mouse, produced airway epithelium with normal histology [38].…”

Section: Discussionmentioning

confidence: 99%

Generation of a human airway epithelium derived basal cell line with multipotent differentiation capacity

et al. 2013

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BackgroundAs the multipotent progenitor population of the airway epithelium, human airway basal cells (BC) replenish the specialized differentiated cell populations of the mucociliated airway epithelium during physiological turnover and repair. Cultured primary BC divide a limited number of times before entering a state of replicative senescence, preventing the establishment of long-term replicating cultures of airway BC that maintain their original phenotype.MethodsTo generate an immortalized human airway BC cell line, primary human airway BC obtained by brushing the airway epithelium of healthy nonsmokers were infected with a retrovirus expressing human telomerase (hTERT). The resulting immortalized cell line was then characterized under non-differentiating and differentiating air-liquid interface (ALI) culture conditions using ELISA, TaqMan quantitative PCR, Western analysis, and immunofluorescent and immunohistochemical staining analysis for cell type specific markers. In addition, the ability of the cell line to respond to environmental stimuli under differentiating ALI culture was assessed.ResultsWe successfully generated an immortalized human airway BC cell line termed BCi-NS1 via expression of hTERT. A single cell derived clone from the parental BCi-NS1 cells, BCi-NS1.1, retains characteristics of the original primary cells for over 40 passages and demonstrates a multipotent differentiation capacity into secretory (MUC5AC, MUC5B), goblet (TFF3), Clara (CC10) and ciliated (DNAI1, FOXJ1) cells on ALI culture. The cells can respond to external stimuli such as IL-13, resulting in alteration of the normal differentiation process.ConclusionDevelopment of immortalized human airway BC that retain multipotent differentiation capacity over long-term culture should be useful in understanding the biology of BC, the response of BC to environmental stress, and as a target for assessment of pharmacologic agents.

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Drug Delivery Characteristics of the Progenitor Bronchial Epithelial Cell Line VA10

Cited by 12 publications

References 47 publications

Investigation on Ciliary Functionality of Different Airway Epithelial Cell Lines in Three-Dimensional Cell Culture

Investigation on Ciliary Functionality of Different Airway Epithelial Cell Lines in Three-Dimensional Cell Culture

Bordetella pertussis Adenylate Cyclase Toxin Disrupts Functional Integrity of Bronchial Epithelial Layers

Generation of a human airway epithelium derived basal cell line with multipotent differentiation capacity

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