Growth and differentiation of primary and passaged equine bronchial epithelial cells under conventional and air-liquid-interface culture conditions

Abraham, Getu; Zizzadoro, Claudia; Kacza, Johannes; Ellenberger, C.; Abs, Vanessa; Franke, Jana; Schoon, Heinz-Adolf; Seeger, Johannes; Tesfaigzi, Yohannes; Ungemach, F. R.

doi:10.1186/1746-6148-7-26

Cited by 33 publications

(44 citation statements)

References 36 publications

(62 reference statements)

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“…EBF were cultured under the indicated culture conditions on glass cover slips (Carl Roth GmbH, Karlsruhe, Germany), washed twice with PBS after medium removal, and fixed in ice-cold acetone for 5 min at -20°C and immunostained as previously described [36]. …”

Section: Methodsmentioning

confidence: 99%

Comparative study of the effects of fetal bovine serum versus horse serum on growth and differentiation of primary equine bronchial fibroblasts

et al. 2014

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BackgroundAirway fibroblasts have become a critical addition to all facets of structural lung tissue changes such as in human asthma and chronic obstructive pulmonary disease, but little is known about their role in the equine recurrent airway obstruction, a disease that resembles to the human asthma. Since the equine bronchial fibroblasts (EBF) have not been isolated and characterized yet, the use of defined medium was investigated.ResultsPrimary EBF were cultured on non-collagen coated flasks without serum or in the presence of fetal bovine serum (FBS) or horse serum (HS) or in serum depleted medium. EBF cultured in serum-free basal media and those serum deprived were not able to proliferate and even exhibited considerable cell death. In media containing FBS or HS, proliferation of the cells was reproducible between different primary cultures and cells demonstrated expression of vimentin. Large variations were found in the ability of FBS and HS to support growth and differentiation of EBF in monolayer culture. Indications of growth-promoting actions, increasing passage number as well as maintaining fibroblast morphology were found rather in FBS than in HS. EBF culturing in HS needed longer doubling and confluence time. The protein content of the cell pellets was higher in EBF cultured in medium containing HS than FBS. Alpha-smooth muscle actin seemed to be less expressed in EBF cultured in medium containing FBS than those in HS.ConclusionsIn sum, serum addition to basal EBF medium enhanced EBF differentiation into myofibroblasts, and these findings are useful to develop in vitro fibroblast culture models that mimic in vivo physiological processes and to study airway disease mechanisms and remodeling.

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Section: Methodsmentioning

confidence: 99%

Comparative study of the effects of fetal bovine serum versus horse serum on growth and differentiation of primary equine bronchial fibroblasts

et al. 2014

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“…Successful cultures of primary and immortalized cells of airway epithelial origin are already well established for humans and a few animal species. These include mainly equine, porcine, bovine, and mouse cell lines [22][23][24][25], but to the best of our knowledge, no characterized airway epithelial cell lines from bats exist.…”

Section: Discussionmentioning

confidence: 99%

Bat Airway Epithelial Cells: A Novel Tool for the Study of Zoonotic Viruses

et al. 2014

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Bats have been increasingly recognized as reservoir of important zoonotic viruses. However, until now many attempts to isolate bat-borne viruses in cell culture have been unsuccessful. Further, experimental studies on reservoir host species have been limited by the difficulty of rearing these species. The epithelium of the respiratory tract plays a central role during airborne transmission, as it is the first tissue encountered by viral particles. Although several cell lines from bats were established recently, no well-characterized, selectively cultured airway epithelial cells were available so far. Here, primary cells and immortalized cell lines from bats of the two important suborders Yangochiroptera and Yinpterochiroptera, Carollia perspicillata (Seba's short-tailed bat) and Eidolon helvum (Straw-colored fruit bat), were successfully cultured under standardized conditions from both fresh and frozen organ specimens by cell outgrowth of organ explants and by the use of serum-free primary cell culture medium. Cells were immortalized to generate permanent cell lines. Cells were characterized for their epithelial properties such as expression of cytokeratin and tight junctions proteins and permissiveness for viral infection with Rift-Valley fever virus and vesicular stomatitis virus Indiana. These cells can serve as suitable models for the study of bat-borne viruses and complement cell culture models for virus infection in human airway epithelial cells.

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“…The TEER is an important method for assessing the formation of junctional complexes between cells [54]. In our model, once confluency was reached, the TEER of the culture rapidly increased (Fig 3C) [25,44,55], however TEER of other AECMs have been shown to stabilised at the peak [32]. This variation in TEER over the course of culturing was not reflected in the presence of tight-junctional protein ZO-1, which was maintained at a stable level throughout the time-course.…”

Section: The Formation Of Tight-junctions Between Cells Creates a Phymentioning

confidence: 91%

Temporal differentiation of bovine airway epithelial cells grown at an air-liquid interface

Cozens

Sutherland

Marchesi

et al. 2017

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17The respiratory epithelium is exposed to assault by toxins and pathogens through the process 18 of inhalation, which has numerous implications on both human and animal health. As such, 19 there is a need to develop and characterise an in vitro model of the airway epithelium to study 20 respiratory pathologies during infection or toxicology experiments. This has been achieved 21 by growing airway epithelial cells at an air-liquid interface (ALI). Characterisation of ALI 22 models are not well-defined for airway epithelial cells derived from non-human species. In 23 this study we have fully characterised a bovine airway epithelial cell models (AECM) grown 24 at an ALI in relation to ex vivo tissue. The morphology of the model was monitored at three 25 day intervals, to identify the time-period at which the culture was optimally differentiated. derived from donor animals, our bovine AECM was shown to be highly representative of the 35 in vivo bovine bronchial epithelium and can be utilised in the study of respiratory 36 pathologies. 37 peer-reviewed)

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Growth and differentiation of primary and passaged equine bronchial epithelial cells under conventional and air-liquid-interface culture conditions

Cited by 33 publications

References 36 publications

Comparative study of the effects of fetal bovine serum versus horse serum on growth and differentiation of primary equine bronchial fibroblasts

Comparative study of the effects of fetal bovine serum versus horse serum on growth and differentiation of primary equine bronchial fibroblasts

Bat Airway Epithelial Cells: A Novel Tool for the Study of Zoonotic Viruses

Temporal differentiation of bovine airway epithelial cells grown at an air-liquid interface

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